Patent Publication Number: US-2009220991-A1

Title: Reagents for the detection of protein phosphorylation in leukemia signaling pathways

Description:
RELATED APPLICATIONS 
     This application claims the benefit of, and priority to, PCT serial number PCT/US06/034126, filed Aug. 30, 2006, presently pending, the disclosure of which is incorporated herein, in its entirety, by reference. 
    
    
     FIELD OF THE INVENTION 
     The invention relates generally to antibodies and peptide reagents for the detection of protein phosphorylation, and to protein phosphorylation in cancer. 
     BACKGROUND OF THE INVENTION 
     The activation of proteins by post-translational modification is an important cellular mechanism for regulating most aspects of biological organization and control, including growth, development, homeostasis, and cellular communication. Protein phosphorylation, for example, plays a critical role in the etiology of many pathological conditions and diseases, including cancer, developmental disorders, autoimmune diseases, and diabetes. Yet, in spite of the importance of protein modification, it is not yet well understood at the molecular level, due to the extraordinary complexity of signaling pathways, and the slow development of technology necessary to unravel it. 
     Protein phosphorylation on a proteome-wide scale is extremely complex as a result of three factors: the large number of modifying proteins, e.g. kinases, encoded in the genome, the much larger number of sites on substrate proteins that are modified by these enzymes, and the dynamic nature of protein expression during growth, development, disease states, and aging. The human genome, for example, encodes over 520 different protein kinases, making them the most abundant class of enzymes known. See Hunter,  Nature  411: 355-65 (2001). Most kinases phosphorylate many different substrate proteins, at distinct tyrosine, serine, and/or threonine residues. Indeed, it is estimated that one-third of all proteins encoded by the human genome are phosphorylated, and many are phosphorylated at multiple sites by different kinases. See Graves et al.,  Pharmacol. Ther.  82:111-21 (1999). 
     Many of these phosphorylation sites regulate critical biological processes and may prove to be important diagnostic or therapeutic targets for molecular medicine. For example, of the more than 100 dominant oncogenes identified to date, 46 are protein kinases. See Hunter, supra. Understanding which proteins are modified by these kinases will greatly expand our understanding of the molecular mechanisms underlying oncogenic transformation. Therefore, the identification of, and ability to detect, phosphorylation sites on a wide variety of cellular proteins is crucially important to understanding the key signaling proteins and pathways implicated in the progression of diseases like cancer. 
     One form of cancer in which underlying signal transduction events are involved, but still poorly understood, is leukemia. Leukemia is a malignant disease of the bone marrow and blood, characterized by abnormal accumulation of blood cells, and is divided in four major categories. An estimated 33,500 new cases of leukemia will be diagnosed in the U.S. alone this year, affecting roughly 30,000 adults and 3,000 children, and close to 24,000 patients will die from the disease (Source: The Leukemia &amp; Lymphoma Society (2004)). Depending of the cell type involved and the rate by which the disease progresses it can be defined as acute or chronic myelogenous leukemia (AML or CML), or acute and chronic lymphocytic leukemia (ALL or CLL). The acute forms of the disease rapidly progress, causing the accumulation of immature, functionless cells in the marrow and blood, which in turn results in anemia, immunodeficiency and coagulation deficiencies, respectively. Chronic forms of leukemia progress more slowly, allowing a greater number of mature, functional cells to be produced, which amass to high concentration in the blood over time. 
     More than half of adult leukemias occur in patients 67 years of age or older, and leukemia accounts for about 30% of all childhood cancers. The most common type of adult leukemia is acute myelogenous leukemia (AML), with an estimated 11,920 new cases annually. Without treatment patients rarely survive beyond 6-12 months, and despite continued development of new therapies, it remains fatal in 80% of treated patients (Source: The Leukemia &amp; Lymphoma Society (2004)). The most common childhood leukemia is acute lymphocytic leukemia (ALL), but it can develop at any age. Chronic lymphocytic leukemia (CLL) is the second most prevalent adult leukemia, with approximately 8,200 new cases of CLL diagnosed annually in the U.S. The course of the disease is typically slower than acute forms, with a five-year relative survival of 74%. Chronic myelogenous leukemia (CML) is less prevalent, with about 4,600 new cases diagnosed each year in the U.S., and is rarely observed in children. 
     Most varieties of leukemia are generally characterized by genetic alterations associated with the etiology of the disease, and it has recently become apparent that, in many instances, such alterations (chromosomal translocations, deletions or point mutations) result in the constitutive activation of protein kinase genes, and their products, particularly tyrosine kinases. The most well known alteration is the oncogenic role of the chimeric BCR-Abl gene, which is generated by translocation of chromosome 9 to chromosome 22, creating the so-called Philadelphia chromosome characteristic of CML (see Nowell,  Science  132: 1497 (1960)). The resulting BCR-Abl kinase protein is constitutively active and elicits characteristic signaling pathways that have been shown to drive the proliferation and survival of CML cells (see Daley,  Science  247: 824-830 (1990); Raitano et al.,  Biochim. Biophys. Acta. December  9; 1333(3): F201-16 (1997)). The recent success of Imanitib (also known as ST1571 or Gleevec®), the first molecularly targeted compound designed to specifically inhibit the tyrosine kinase activity of BCR-Abl, provided critical confirmation of the central role of BCR-Abl signaling in the progression of CML (see Schindler et al.,  Science  289: 1938-1942 (2000); Nardi et al.,  Curr. Opin. Hematol.  11: 35-43 (2003)). 
     The success of Gleevec® now serves as a paradigm for the development of targeted drugs designed to block the activity of other tyrosine kinases known to be involved in leukemias and other malignancies (see, e.g., Sawyers,  Curr. Opin. Genet. Dev. February;  12(1): 111-5 (2002); Druker,  Adv. Cancer Res.  91:1-30 (2004)). For example, recent studies have demonstrated that mutations in the FLT3 gene occur in one third of adult patients with AML. FLT3 (Fms-like tyrosine kinase 3) is a member of the class III receptor tyrosine kinase (RTK) family including FMS, platelet-derived growth factor receptor (PDGFR) and c-KIT (see Rosnet et al.,  Crit. Rev. Oncog.  4: 595-613 (1993). In 20-27% of patients with AML, an internal tandem duplication in the juxta-membrane region of FLT3 can be detected (see Yokota et al.,  Leukemia  11: 1605-1609 (1997)). Another 7% of patients have mutations within the active loop of the second kinase domain, predominantly substitutions of aspartate residue 835 (D835), while additional mutations have been described (see Yamamoto et al.,  Blood  97: 2434-2439 (2001); Abu-Duhier et al.,  Br. J. Haematol.  113: 983-988 (2001)). Expression of mutated FLT3 receptors results in constitutive tyrosine phosphorylation of FLT3, and subsequent phosphorylation and activation of downstream molecules such as STAT5, Akt and MAPK, resulting in factor-independent growth of hematopoietic cell lines. 
     Altogether, FLT3 is the single most common activated gene in AML known to date. This evidence has triggered an intensive search for FLT3 inhibitors for clinical use leading to at least four compounds in advanced stages of clinical development, including: PKC412 (by Novartis), CEP-701 (by Cephalon), MLN518 (by Millenium Pharmaceuticals), and SU5614 (by Sugen/Pfizer) (see Stone et al.,  Blood  (in press)(2004); Smith et al.,  Blood  103: 3669-3676 (2004); Clark et al.,  Blood  104: 2867-2872 (2004); and Spiekerman et al.,  Blood  101: 1494-1504 (2003)). 
     There is also evidence indicating that kinases such as FLT3, c-KIT and Abl are implicated in some cases of ALL (see Cools et al.,  Cancer Res.  64: 6385-6389 (2004); Hu,  Nat. Genet.  36:453461 (2004); and Graux et al.,  Nat. Genet.  36: 1084-1089 (2004)). In contrast, very little is know regarding any causative role of protein kinases in CLL, except for a high correlation between high expression of the tyrosine kinase ZAP70 and the more aggressive form of the disease (see Rassenti et al.,  N. Eng. J. Med.  351: 893-901 (2004)). 
     Despite the identification of a few key molecules involved in progression of leukemia, the vast majority of signaling protein changes underlying this disease remains unknown. There is, therefore, relatively scarce information about kinase-driven signaling pathways and phosphorylation sites relevant to the different types of leukemia. This has hampered a complete and accurate understanding of how protein activation within signaling pathways is driving these complex cancers. Accordingly, there is a continuing and pressing need to unravel the molecular mechanisms of kinase-driven oncogenesis in leukemia by identifying the downstream signaling proteins mediating cellular transformation in this disease. Identifying particular phosphorylation sites on such signaling proteins and providing new reagents, such as phospho-specific antibodies and AQUA peptides, to detect and quantify them remains particularly important to advancing our understanding of the biology of this disease. 
     Presently, diagnosis of leukemia is made by tissue biopsy and detection of different cell surface markers. However, misdiagnosis can occur since some leukemia cases can be negative for certain markers, and because these markers may not indicate which genes or protein kinases may be deregulated. Although the genetic translocations and/or mutations characteristic of a particular form of leukemia can be sometimes detected, it is clear that other downstream effectors of constitutively active kinases having potential diagnostic, predictive, or therapeutic value, remain to be elucidated. Accordingly, identification of downstream signaling molecules and phosphorylation sites involved in different types of leukemia and development of new reagents to detect and quantify these sites and proteins may lead to improved diagnostic/prognostic markers, as well as novel drug targets, for the detection and treatment of this disease. 
     SUMMARY OF THE INVENTION 
     The invention discloses nearly 480 novel phosphorylation sites identified in signal transduction proteins and pathways underlying human Leukemias and provides new reagents, including phosphorylation-site specific antibodies and AQUA peptides, for the selective detection and quantification of these phosphorylated sites/proteins. Also provided are methods of using the reagents of the invention for the detection, quantification, and profiling of the disclosed phosphorylation sites. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       FIG.  1 —Is a diagram broadly depicting the immunoaffinity isolation and mass-spectrometric characterization methodology (IAP) employed to identify the novel phosphorylation sites disclosed herein. 
       FIG.  2 —Is a table (corresponding to Table 1) enumerating the Leukemia signaling protein phosphorylation sites disclosed herein: Column A=the name of the parent protein; Column B=the SwissProt accession number for the protein (human sequence); Column C=the protein type/classification; Column D=the tyrosine or serine residue (in the parent protein amino acid sequence) at which phosphorylation occurs within the phosphorylation site; Column E=the phosphorylation site sequence encompassing the phosphorylatable residue (residue at which phosphorylation occurs (and corresponding to the respective entry in Column D) appears in lowercase; Column F=the type of leukemia in which the phosphorylation site was discovered; and Column G=the cell type(s), tissue(s) and/or patient tissue(s) in which the phosphorylation site was discovered. 
       FIG.  3 —is an exemplary mass spectrograph depicting the detection of the tyrosine 330 phosphorylation site in DOK2 (see Row 24 in FIG.  2 /Table 1), as further described in Example 1 (red and blue indicate ions detected in MS/MS spectrum); Y* indicates the phosphorylated tyrosine (shown as lowercase “y” in  FIG. 2 ). 
       FIG.  4 —is an exemplary mass spectrograph depicting the detection of the tyrosine 630 phosphorylation site in FLT3 (see Row 286 in FIG.  2 /Table 1), as further described in Example 1 (red and blue indicate ions detected in MS/MS spectrum); Y* indicates the phosphorylated tyrosine (shown as lowercase “y” in  FIG. 2 ). 
       FIG.  5 —is an exemplary mass spectrograph depicting the detection of the tyrosine 1736 phosphorylation site in TSC2 (see Row 87 in FIG.  2 /Table 1), as further described in Example 1 (red and blue indicate ions detected in MS/MS spectrum); Y* indicates the phosphorylated tyrosine (shown as lowercase “y” in  FIG. 2 ). 
       FIG.  6 —is an exemplary mass spectrograph depicting the detection of the tyrosine 260 phosphorylation site in ICAM (see Row 71 in FIG.  2 /Table 1), as further described in Example 1 (red and blue indicate ions detected in MS/MS spectrum); Y* indicates the phosphorylated tyrosine (shown as lowercase “y” in  FIG. 2 ). 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     In accordance with the present invention, nearly 480 novel protein phosphorylation sites in signaling proteins and pathways underlying human Leukemia have now been discovered. These newly described phosphorylation sites were identified by employing the techniques described in “Immunoaffinity Isolation of Modified Peptides From Complex Mixtures,” U.S. Patent Publication No. 20030044848, Rush et al., using cellular extracts from a variety of leukemia-derived cell lines, e.g. SEM, HT-93, etc., as further described below. The novel phosphorylation sites (tyrosine), and their corresponding parent proteins, disclosed herein are listed in Table 1. These phosphorylation sites correspond to numerous different parent proteins (the full sequences of which (human) are all publicly available in SwissProt database and their Accession numbers listed in Column B of Table 1/ FIG. 2 ), each of which fall into discrete protein type groups, for example Adaptor/Scaffold proteins, Cytoskeletal proteins, Protein Kinases, and Adhesion proteins, etc. (see Column C of Table 1), the phosphorylation of which is relevant to signal transduction activity underlying Leukemias (AML, CML, CLL, and ALL), as disclosed herein. 
     The discovery of the nearly 480 novel protein phosphorylation sites described herein enables the production, by standard methods, of new reagents, such as phosphorylation site-specific antibodies and AQUA peptides (heavy-isotope labeled peptides), capable of specifically detecting and/or quantifying these phosphorylated sites/proteins. Such reagents are highly useful, inter alia, for studying signal transduction events underlying the progression of Leukemia. Accordingly, the invention provides novel reagents—phospho-specific antibodies and AQUA peptides—for the specific detection and/or quantification of a Leukemia-related signaling protein/polypeptide only when phosphorylated (or only when not phosphorylated) at a particular phosphorylation site disclosed herein. The invention also provides methods of detecting and/or quantifying one or more phosphorylated Leukemia-related signaling proteins using the phosphorylation-site specific antibodies and AQUA peptides of the invention, and methods of obtaining a phosphorylation profile of such proteins (e.g. Kinases). 
     In part, the invention provides an isolated phosphorylation site-specific antibody that specifically binds a given Leukemia-related signaling protein only when phosphorylated (or not phosphorylated, respectively) at a particular tyrosine enumerated in Column D of Table 1/ FIG. 2  comprised within the phosphorylatable peptide site sequence enumerated in corresponding Column E. In further part, the invention provides a heavy-isotope labeled peptide (AQUA peptide) for the detection and quantification of a given Leukemia-related signaling protein, the labeled peptide comprising a particular phosphorylatable peptide site/sequence enumerated in Column E of Table 1/ FIG. 2  herein. For example, among the reagents provided by the invention is an isolated phosphorylation site-specific antibody that specifically binds the MELK tyrosine kinase only when phosphorylated (or only when not phosphorylated) at tyrosine 438 (see Row 244 (and Columns D and E) of Table 1/ FIG. 2 ). By way of further example, among the group of reagents provided by the invention is an AQUA peptide for the quantification of phosphorylated MELK tyrosine kinase, the AQUA peptide comprising the phosphorylatable peptide sequence listed in Column E, Row 244, of Table 1/ FIG. 2  (which encompasses the phosphorylatable tyrosine at position 438). 
     In one embodiment, the invention provides an isolated phosphorylation site-specific antibody that specifically binds a human Leukemia-related signaling protein selected from Column A of Table 1 (Rows 2-481) only when phosphorylated at the tyrosine residue listed in corresponding Column D of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E of Table 1 (SEQ ID NOs: 1-7, 9-14, 16, 19-21, 23, 26-30, 32-34, 36-45, 48-52, 56-58, 60-90, 93-119, 121-124, 129-151, 153-160, 163-180, 182-193, 195-197, 199-208, 210-221, 223-279, 281-294, 296-297, 299-316, 319-336, 339-345, 347-356, 358, 360-366, 368-378, 380-417, 419-438, 440-474, and 476-480), wherein said antibody does not bind said signaling protein when not phosphorylated at said tyrosine. In another embodiment, the invention provides an isolated phosphorylation site-specific antibody that specifically binds a Leukemia-related signaling protein selected from Column A of Table 1 only when not phosphorylated at the tyrosine residue listed in corresponding Column D of Table 1, comprised within the peptide sequence listed in corresponding Column E of Table 1 (SEQ ID NOs: 1-7, 9-14, 16, 19-21, 23, 26-30, 32-34, 36-45, 48-52, 56-58, 60-90, 93-119, 121-124, 129-151, 153-160, 163-180, 182-193, 195-197, 199-208, 210-221, 223-279, 281-294, 296-297, 299-316, 319-336, 339-345, 347-356, 358, 360-366, 368-378, 380-417, 419-438, 440-474, and 476-480), wherein said antibody does not bind said signaling protein when phosphorylated at said tyrosine. Such reagents enable the specific detection of phosphorylation (or non-phosphorylation) of a novel phosphorylatable site disclosed herein. The invention further provides immortalized cell lines producing such antibodies. In one preferred embodiment, the immortalized cell line is a rabbit or mouse hybridoma. 
     In another embodiment, the invention provides a heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein selected from Column A of Table 1, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E of Table 1 (SEQ ID NOs: 1-7, 9-14, 16, 19-21, 23, 26-30, 32-34, 36-45, 48-52, 56-58, 60-90, 93-119, 121-124, 129-151, 153-160, 163-180, 182-193, 195-197, 199-208, 210-221, 223-279, 281-294, 296-297, 299-316, 319-336, 339-345, 347-356, 358, 360-366,-368-378, 380-417, 419-438, 440-474, and 476-480), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D of Table 1. In certain preferred embodiments, the phosphorylatable tyrosine within the labeled peptide is phosphorylated, while in other preferred embodiments, the phosphorylatable residue within the labeled peptide is not phosphorylated. 
     Reagents (antibodies and AQUA peptides) provided by the invention may conveniently be grouped by the type of Leukemia-related signaling protein in which a given phosphorylation site (for which reagents are provided) occurs. The protein types for each respective protein (in which a phosphorylation site has been discovered) are provided in Column C of Table 1/ FIG. 2 , and include: adaptor/scaffold proteins, acetyltransferases, actin binding proteins, adhesion proteins, apoptosis proteins, calcium-binding proteins, cell cycle regulation proteins, cell surface proteins, channel proteins, chaperone proteins, contractile proteins, cytokine proteins, cytoskeletal proteins, G protein regulators and GTPase activating proteins, guanine nucleotide exchange factors, helicase proteins, immunoglobulin superfamily proteins, inhibitor proteins, protein kinases, lipid kinases, ligases, lipid binding proteins, methytransferases, motor proteins, oxidoreductases, phosphotases, phosphodiesterases, phospholipases, proteases, receptor proteins, transcription factors, transferases, translation/transporter proteins, and ubiquitin conjugating system proteins. Each of these distinct protein groups is considered a preferred subset of Leukemia-related signal transduction protein phosphorylation sites disclosed herein, and reagents for their detection/quantification may be considered a preferred subset of reagents provided by the invention. 
     Particularly preferred subsets of the phosphorylation sites (and their corresponding proteins) disclosed herein are those occurring on the following protein types/groups listed in Column C of Table 1/ FIG. 2 , are the protein kinases adaptor/scaffold proteins, adhesion proteins, cell cycle regulation proteins, cell surface proteins, transcription proteins, phosphatases, phospholipases, phosphodiesterases, receptor proteins, cytoskeltal proteins, G protein regulators, and lipid kinases. Accordingly, among preferred subsets of reagents provided by the invention are isolated antibodies and AQUA peptides useful for the detection and/or quantification of the foregoing preferred protein/phosphorylation site subsets. 
     In one subset of preferred embodiments, there is provided:
         (i) An isolated phosphorylation site-specific antibody that specifically binds a protein kinase selected from Column A, Rows 210-291, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 210-291, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 210-291, of Table 1 (SEQ ID NOs: 210-221, 223-280, and 281-290), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.       (ii) An equivalent antibody to (i) above that only binds the protein kinase when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a protein kinase selected from Column A, Rows 210-291, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 210-291, of Table 1 (SEQ ID NOs: 210-221, 223-280, and 281-290), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 210-291, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following protein kinase phosphorylation sites are particularly preferred: ATM (Y2129), MELK (Y438), MAPK14 (Y24), BLK (Y187), BTK (Y344), SYK (Y296), ZAP70 (Y69), FLT3 (Y630), FLT3 (Y726), FLT3 (Y768), and ROS1 (Y363) (see SEQ ID NOs: 229, 243, 249, 270-271, 273, 283, and 285-288). 
     In a second subset of preferred embodiments there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds an adaptor/scaffold protein selected from Column A, Rows 11-59, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 11-59, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 11-59, of Table 1 (SEQ ID NOs: 10-14, 16,19-21, 23, 26-30, 32-34, 36-45, 48-52, and 56-58), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the adaptor/scaffold protein when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is a adaptor/scaffold protein selected from Column A, Rows 11-59, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 11-59, of Table 1 (SEQ ID NOs: 10-14, 16, 19-21, 23, 26-30, 32-34, 36-45, 48-52, and 56-58), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 11-59, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following adaptor/scaffold protein phosphorylation sites are particularly preferred: ABI2 (Y192), PIK3AP1 (Y594), DOK2 (Y330), LAT2 (Y40), SIT1 (Y127), STAM (Y384), SCAP1 (Y142) (see SEQ ID NOs: 10,14, 23, 30, 37, 40-41). 
     In another subset of preferred embodiments there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds an adhesion protein selected from Column A, Rows 60-79, of Table 1 only when phosphorylated at the tyrosine or serine listed in corresponding Column D, Rows 60-79, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 60-79, of Table 1 (SEQ ID NOs: 60-78), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the adhesion protein when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is an adhesion protein selected from Column A, Rows 60-79, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 60-79, of Table 1 (SEQ ID NOs: 60-78), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 60-79, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following adhesion protein phosphorylation sites are particularly preferred: ADAM18 (Y197), ICAM2 (Y260) and PECAM1 (Y663) (see SEQ ID NOs: 60, 70 and 72). 
     In still another subset of preferred embodiments there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds a cell cycle regulation protein selected from Column A, Rows 83-87, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 83-87, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 83-87, of Table 1 (SEQ ID NOs: 82-86), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the cell cycle regulation protein when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is a cell cycle regulation protein selected from Column A, Rows 83-87, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 83-87, of Table 1 (SEQ ID NOs: 82-86), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 83-87, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following cell cycle regulation protein phosphorylation sites are particularly preferred: TSC2 (Y1736) (see SEQ ID NO: 86). 
     In still another subset of preferred embodiments there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds a cell surface protein selected from Column A, Rows 88-94, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 88-94, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 88-94, of Table 1 (SEQ ID NOs: 87-90, and 93), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the cell surface protein when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is a cell surface protein selected from Column A, Rows 88-94, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 88-94, of Table 1 (SEQ ID NOs: 87-90, and 93), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 88-94, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following cell surface protein phosphorylation sites are particularly preferred: CD72 (Y39) and CD84 (Y299) (see SEQ ID NOs: 89 and 93). 
     In still another subset of preferred embodiments there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds a transcription factor/coactivator/corepressor selected from Column A, Rows 387-425, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 387-425, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 387-425 of Table 1 (SEQ ID NOs: 386-417, and 419-424), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds transcription factor/coactivator/corepressor when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is a transcription factor/coactivator/corepressor selected from Column A, Rows 387-425, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 387-425, of Table 1 (SEQ ID NOs: 386-417, and 419-424), which sequence comprises the phosphorylatable tyrosine or serine listed in corresponding Column D, Rows 387-425, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following trascription factor/coactivor/corepressor phosphorylation sites are particularly preferred: STAT5A (Y22), STAT5A (Y90), STAT5A (Y1 14), SMAD2 (Y102), and NSEP1 (Y208) (see SEQ ID NOs: 410-413 and 417). 
     In yet another subset of preferred embodiments, there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds a phophatase selected from Column A, Rows 331-350, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 331-350, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 331-350, of Table 1 (SEQ ID NOs: 330-349), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the phosphatase when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is a phophatase selected from Column A, Rows 331-350, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 331-350, of Table 1 (SEQ ID NOs: 330-349), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 331-350, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following phosphatase phosphorylation sites are particularly preferred: INPPL1 (Y831), INPPL1 (Y1135) and PTPRC (Y705) (see SEQ ID NOs: 336-337, and 347). 
     In yet another subset of preferred embodiments, there is provided:
     (i) An isolated phosphorylation site-specific antibody specifically binds a phosphodiesterase/phospholipase selected from Column A, Rows 351-359, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 351-359, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 351-359, of Table 1 (SEQ ID NOs: 350-356, and 358), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the Tyrosine Protein Kinase when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is a phosphodiesterase/phospholipase selected from Column A, Rows 351-359, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 351-359, of Table 1 (SEQ ID NOs: 350-356, and 358), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 351-359, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following phosphodiesterase/phospholipase phosphorylation sites are particularly preferred: PLCG1 (Y481), PLCG2 (Y680) and PLCG2 (Y1264) (see SEQ ID NOs: 352, 354 and 358). 
     In yet another subset of preferred embodiments, there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds a receptor protein selected from Column A, Rows 366-386, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 366-386, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 366-386, of Table 1 (SEQ ID NOs: 365-366, 368-378, and 380-385), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the receptor protein when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is a receptor protein selected from Column A, Rows 366-386, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 366-386, of Table 1 (SEQ ID NOs: 365-366, 368-378, and 380-385), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 366-386, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following receptor protein phosphorylation sites are particularly preferred: LEPR (Y795) (see SEQ ID NO: 365). 
     In still another subset of preferred embodiments, there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds a cytoskeletal protein selected from Column A, Rows 114-170, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 114-170, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 114-170, of Table 1 (SEQ ID NOs: 113-119, 121-124, 129-151, 153-160, and 163-169), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the cytoskeletal protein when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that cytoskeletal protein selected from Column A, Rows 114-170, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 114-170, of Table 1 (SEQ ID NOs: 113-119, 121-124, 129-151, 153-160, and 163-169), which sequence comprises the phosphorylatable tyrosine or serine listed in corresponding Column D, Rows 114-170, of Table 1.   

     In still another subset of preferred embodiments, there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds a G protein selected from Column A, Rows 174-180, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 174-180, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 174-180, of Table 1 (SEQ ID NOs: 173-189), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the G protein when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is an G protein selected from Column A, Rows 174-180, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 174-180, of Table 1 (SEQ ID NOs: 173-179), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 174-180, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following G protein phosphorylation sites are particularly preferred: GNAI2 (Y61) (see SEQ ID NO: 177). 
     In still another subset of preferred embodiments, there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds a lipid kinase selected from Column A, Rows 292-298, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 292-298, of Table 1, comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 292-298, of Table 1 (SEQ ID NOs: 291-297), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the lipid kinase when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a Leukemia-related signaling protein that is an lipid kinase selected from Column A, Rows 292-298, said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 292-298, of Table 1 (SEQ ID NOs: 291-297), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 292-298, of Table 1.   

     Among this preferred subset of reagents, antibodies and AQUA peptides for the detection/quantification of the following lipid kinase phosphorylation sites are particularly preferred: PIK3CB (Y962) (see SEQ ID NO: 293). 
     In yet a further subset of preferred embodiments, there is provided:
     (i) An isolated phosphorylation site-specific antibody that specifically binds a protein selected from the group consisting of EIF4EBP2, EIF4G2 and EIF4B (Column A, Rows 446, 448 and 460 of Table 1) only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 191, 199, 446, 448 and 460 of Table 1), said tyrosine comprised within the phosphorylatable peptide sequence listed in corresponding Column E, Rows 446, 448 and 460, of Table 1 (SEQ ID NOs: 445, 447 and 459), wherein said antibody does not bind said protein when not phosphorylated at said tyrosine.   (ii) An equivalent antibody to (i) above that only binds the EIF4EBP2, EIF4G2 and EIF4B protein when not phosphorylated at the disclosed site (and does not bind the protein when it is phosphorylated at the site).   (iii) A heavy-isotope labeled peptide (AQUA peptide) for the quantification of a protein selected from the group consisting of EIF4EBP2, EIF4G2 and EIF4B (Column A, Rows 446, 448 and 460 of Table 1), said labeled peptide comprising the phosphorylatable peptide sequence listed in corresponding Column E, Rows 446, 448 and 460, of Table 1 (SEQ ID NOs: 445, 447 and 459), which sequence comprises the phosphorylatable tyrosine listed in corresponding Column D, Rows 446, 448 and 460, of Table 1.   

     The invention also provides, in part, an immortalized cell line producing an antibody of the invention, for example, a cell line producing an antibody within any of the foregoing preferred subsets of antibodies. In one preferred embodiment, the immortalized cell line is a rabbit hybridoma or a mouse hybridoma. 
     In certain other preferred embodiments, a heavy-isotope labeled peptide (AQUA peptide) of the invention (for example, an AQUA peptide within any of the foregoing preferred subsets of AQUA peptides) comprises a disclosed site sequence wherein the phosphorylatable tyrosine is phosphorylated. In certain other preferred embodiments, a heavy-isotope labeled peptide of the invention comprises a disclosed site sequence wherein the phosphorylatable tyrosine is not phosphorylated. 
     The foregoing subsets of preferred reagents of the invention should not be construed as limiting the scope of the invention, which, as noted above, includes reagents for the detection and/or quantification of disclosed phosphorylation sites on any of the other protein type/group subsets (each a preferred subset) listed in Column C of Table 1/ FIG. 2 . 
     Also provided by the invention are methods for detecting or quantifying a Leukemia-related signaling protein that is tyrosine phosphorylated, said method comprising the step of utilizing one or more of the above-described reagents of the invention to detect or quantify one or more Leukemia-related signaling protein(s) selected from Column A of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D of Table 1. In certain preferred embodiments of the methods of the invention, the reagents comprise a subset of preferred reagents as described above. 
     Also provided by the invention is a method for obtaining a phosphorylation profile of protein kinases that are phosphorylated in Leukemia signaling pathways, said method comprising the step of utilizing one or more isolated antibody that specifically binds a protein inase selected from Column A, Rows 210-291, of Table 1 only when phosphorylated at the tyrosine listed in corresponding Column D, Rows 210-291, of Table 1, comprised within the phosphorylation site sequence listed in corresponding Column E, Rows 210-291, of Table 1 (SEQ ID NOs: SEQ ID NOs: 210-221, 223-280, and 281-290), to detect the phosphorylation of one or more of said protein kinases, thereby obtaining a phosphorylation profile for said kinases. 
     The identification of the disclosed novel Leukemia-related signaling protein phosphorylation sites, and the standard production and use of the reagents provided by the invention are described in further detail below and in the Examples that follow. 
     All cited references are hereby incorporated herein, in their entirety, by reference. The Examples are provided to further illustrate the invention, and do not in any way limit its scope, except as provided in the claims appended hereto. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Newly Discovered Leukemia-related Phosphorylation Sites. 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                 A 
                   
                   
                   
                   
                   
                   
               
               
                   
                 Protein 
                   
                   
                 D 
                 E 
               
               
                   
                 Name 
                 B 
                 C 
                 Phospho- 
                 Phosphorylation 
                 H 
               
               
                 1 
                 (short) 
                 Accession No. 
                 Protein Type 
                 Residue 
                 Site Sequence 
                 SEQ ID NO 
               
               
                   
               
               
                   2 
                 ZDHHC17 
                 NP_056151.2 
                 Acetyltransferase 
                 Y336 
                 GLMyGGVWATVQFLSKSFFDHSMH 
                 SEQ ID NO: 1 
                   
               
               
                   
                   
                   
                   
                   
                 SALPLGIYLATK 
               
               
                   
               
               
                   3 
                 ZDHHC17 
                 NP_056151.2 
                 Acetyltransferase 
                 Y364 
                 GLMYGGVWATVQFLSKSFFDHSMH 
                 SEQ ID NO: 2 
               
               
                   
                   
                   
                   
                   
                 SALPLGlyLATK 
               
               
                   
               
               
                   4 
                 CNN2 
                 NP_004359.1 
                 Actin binding protein 
                 Y184 
                 CASQSGMTAyGTRR 
                 SEQ ID NO: 3 
               
               
                   
               
               
                   5 
                 DBN1 
                 NP_004386.1 
                 Actin binding protein 
                 Y597 
                 EGTQASEGyFSQSQEEEFAQSEEL 
                 SEQ ID NO: 4 
               
               
                   
                   
                   
                   
                   
                 CAK 
               
               
                   
               
               
                   6 
                 DBNL 
                 NP_001014436.1 
                 Actin binding protein 
                 Y140 
                 VAKASGANySFHK 
                 SEQ ID NO: 5 
               
               
                   
               
               
                   7 
                 FSCN2 
                 NP_036550.1 
                 Actin binding protein 
                 Y352 
                 yVCMKKNGQLAAISDFVGK 
                 SEQ ID NO: 6 
               
               
                   
               
               
                   8 
                 FLNB 
                 NP_001448.2 
                 Actin binding protein 
                 Y414 
                 DIYTAGAGVGDIGVEVEDPQGKNT 
                 SEQ ID NO: 7 
               
               
                   
                   
                   
                   
                   
                 VELLVEDKGNQVy 
               
               
                   
               
               
                   9 
                 LCP1 
                   
                 Actin binding protein 
                 Y28 
                 VDTDGNGyISFNELN 
                 SEQ ID NO: 8 
               
               
                   
               
               
                  10 
                 PIP 
                 NP_002643.1 
                 Actin binding protein 
                 Y71 
                 TyLISSIPLQGAFNYKYTACKCDD 
                 SEQ ID NO: 9 
               
               
                   
                   
                   
                   
                   
                 NPK 
               
               
                   
               
               
                  11 
                 ABI2 
                 NP_005750.4 
                 Adaptor/scaffold 
                 Y192 
                 HSPyRTLEPVRPPVVPNDYVPSPT 
                 SEQ ID NO: 10 
               
               
                   
                   
                   
                   
                   
                 R 
               
               
                   
               
               
                  12 
                 AMOTL1 
                 NP_570899.1 
                 Adaptor/scaffold 
                 Y191 
                 STQPQQNNEELPTyEEAK 
                 SEQ ID NO: 11 
               
               
                   
               
               
                  13 
                 ANK1 
                 NP_000028.3 
                 Adaptor/scaffold 
                 Y1468 
                 EGQNANMENLyTALQSIDRGEIVN 
                 SEQ ID NO: 12 
               
               
                   
                   
                   
                   
                   
                 MLEGSGRQSR 
               
               
                   
               
               
                  14 
                 ARRB2 
                 NP_004304.1 
                 Adaptor/scaffold 
                 Y404 
                 LKGMKDDDyDDQLC 
                 SEQ ID NO: 13 
               
               
                   
               
               
                  15 
                 PIK3AP1 
                 NP_689522.2 
                 Adaptor/scaffold 
                 Y594 
                 DRPQSSIySPFAGMK 
                 SEQ ID NO: 14 
               
               
                   
               
               
                  16 
                 PIK3AP1 
                   
                 Adaptor/scaffold 
                 Y694 
                 AKVEFGVyESGPRKS 
                 SEQ ID NO: 15 
               
               
                   
               
               
                  17 
                 NEDD9 
                 NP_006394.1 
                 Adaptor/scaffold 
                 Y172 
                 yQKDVYDIPPSHTTGQVYDIPPSS 
                 SEQ ID NO: 16 
               
               
                   
                   
                   
                   
                   
                 AK 
               
               
                   
               
               
                  18 
                 NEDD9 
                   
                 Adaptor/scaffold 
                 Y177 
                 YQKDVyDIPPSHTTQGVYDIPPSS 
                 SEQ ID NO: 17 
               
               
                   
                   
                   
                   
                   
                 AK 
               
               
                   
               
               
                  19 
                 NEDD9 
                   
                 Adaptor/scaffold 
                 Y189 
                 DVYDIPPSHTTQGVyDIPPSSAK 
                 SEQ ID NO: 18 
               
               
                   
               
               
                  20 
                 CD2AP 
                 NP_036252.1 
                 Adaptor/scaffold 
                 Y361 
                 yFSLKPEEKDEK 
                 SEQ ID NO: 19 
               
               
                   
               
               
                  21 
                 DIAPH1 
                 NP_005210.2 
                 Adaptor/scaffold 
                 Y415 
                 NDyEARPQYYK 
                 SEQ ID NO: 20 
               
               
                   
               
               
                  22 
                 DAB2 
                 NP_001334.1 
                 Adaptor/scaffold 
                 Y685 
                 KGEQTSSGTLSAFASyFNSK 
                 SEQ ID NO: 21 
               
               
                   
               
               
                  23 
                 DOK2 
                   
                 Adaptor/scaffold 
                 Y139 
                 CMEENELySSAVTVG 
                 SEQ ID NO: 22 
               
               
                   
               
               
                  24 
                 DOK2 
                 NP_003965.2 
                 Adaptor/scaffold 
                 Y330 
                 VPPQLLADPLyDSIEETLPPRPDH 
                 SEQ ID NO: 23 
               
               
                   
                   
                   
                   
                   
                 IYDEPEGV 
               
               
                   
               
               
                  25 
                 DOK2 
                   
                 Adaptor/scaffold 
                 Y345 
                 ADPLYDSIEETLPPRPDHIyDEPE 
                 SEQ ID NO: 24 
               
               
                   
                   
                   
                   
                   
                 GV 
               
               
                   
               
               
                  26 
                 GAB2 
                   
                 Adaptor/scaffold 
                 Y438 
                 AGDNSQSVyIPMSPGAHHFDSLGY 
                 SEQ ID NO: 25 
               
               
                   
                   
                   
                   
                   
                 PSTTLPVHR 
               
               
                   
               
               
                  27 
                 GAB2 
                 NP_036428.1 
                 Adaptor/scaffold 
                 Y525 
                 ANHTFNSSSSQyCR 
                 SEQ ID NO: 26 
               
               
                   
               
               
                  28 
                 C20orf32 
                 NP_065089.2 
                 Adaptor/scaffold 
                 Y312 
                 LSLPEIPSyGFLVPR 
                 SEQ ID NO: 27 
               
               
                   
               
               
                  29 
                 HCLS1 
                 NP_005326.1 
                 Adaptor/scaffold 
                 Y360 
                 GLQVEEEPVyE 
                 SEQ ID NO: 28 
               
               
                   
               
               
                  30 
                 SLC4A1AP 
                 NP_060628.1 
                 Adaptor/scaffold 
                 Y773 
                 SSKYPEDDPDyCVW 
                 SEQ ID NO: 29 
               
               
                   
               
               
                  31 
                 LAT2 
                 NP_054865.2 
                 Adaptor/scaffold 
                 Y40 
                 RSEKIyQQR 
                 SEQ ID NO: 30 
               
               
                   
               
               
                  32 
                 LAT2 
                   
                 Adaptor/scaffold 
                 Y58 
                 SFTGSRTySLVGQAW 
                 SEQ ID NO: 31 
               
               
                   
               
               
                  33 
                 LAT2 
                 NP_054865.2 
                 Adaptor/scaffold 
                 Y84 
                 LLQFyPSLEDPASSR 
                 SEQ ID NO: 32 
               
               
                   
               
               
                  34 
                 PDLIM5 
                 NP_001011513.1 
                 Adaptor/scaffold 
                 Y138 
                 yTEFYHVPTHSDASK 
                 SEQ ID NO: 33 
               
               
                   
               
               
                  35 
                 LIMS1 
                 NP_004978.2 
                 Adaptor/scaffold 
                 Y304 
                 FVEFDMKPVCKKCyEK 
                 SEQ ID NO: 34 
               
               
                   
               
               
                  36 
                 SCAP2 
                   
                 Adaptor/scaffold 
                 Y237 
                 YDERGELyDDVDHPL 
                 SEQ ID NO: 35 
               
               
                   
               
               
                  37 
                 SAMSN1 
                 NP_071419.3 
                 Adaptor/scaffold 
                 Y160 
                 LDDDGPySGPFCGR 
                 SEQ ID NO: 36 
               
               
                   
               
               
                  38 
                 SIT1 
                 NP_055265.1 
                 Adaptor/scaffold 
                 Y127 
                 AAEEVMCyTSLQLRPPQGR 
                 SEQ ID NO: 37 
               
               
                   
               
               
                  39 
                 SIT1 
                 NP_055265.1 
                 Adaptor/scaffold 
                 Y169 
                 SQASGPEPELyASVCAQTR 
                 SEQ ID NO: 38 
               
               
                   
               
               
                  40 
                 STAM 
                 NP_003464.1 
                 Adaptor/scaffold 
                 Y381 
                 LMNEDPMySMYAK 
                 SEQ ID NO: 39 
               
               
                   
               
               
                  41 
                 STAM 
                 NP_003464.1 
                 Adaptor/scaffold 
                 Y384 
                 LMNEDPMYSMyAK 
                 SEQ ID NO: 40 
               
               
                   
               
               
                  42 
                 SCAP1 
                 NP_003717.2 
                 Adaptor/scaffold 
                 Y142 
                 GLFYyYANEK 
                 SEQ ID NO: 41 
               
               
                   
               
               
                  43 
                 FYB 
                 NP_001456.3 
                 Adaptor/scaffold 
                 Y826 
                 YGYVLRSYLADNDGEIYDDIADGC 
                 SEQ ID NO: 42 
               
               
                   
                   
                   
                   
                   
                 IyDND 
               
               
                   
               
               
                  44 
                 SDCBP 
                 NP_001007068.1 
                 Adaptor/scaffold 
                 Y91 
                 PSSINyMVAPVTGNDVGIR 
                 SEQ ID NO: 43 
               
               
                   
               
               
                  45 
                 TRIP6 
                 NP_003293.2 
                 Adaptor/scaffold 
                 Y149 
                 TGSLKPNPASPLPASPyGGPTPAS 
                 SEQ ID NO: 44 
               
               
                   
                   
                   
                   
                   
                 YTTASTPAGPAFPVQVK 
               
               
                   
               
               
                  46 
                 TRIP6 
                 NP_003293.2 
                 Adaptor/scaffold 
                 Y157 
                 TGSLKPNPASPLPASPYGGPTPAS 
                 SEQ ID NO: 45 
               
               
                   
                   
                   
                   
                   
                 yTTASTPAGPAFPVQVK 
               
               
                   
               
               
                  47 
                 TJP2 
                   
                 Adaptor/scaffold 
                 Y423 
                 PEERRHQySDYDYHS 
                 SEQ ID NO: 46 
               
               
                   
               
               
                  48 
                 TJP2 
                   
                 Adaptor/scaffold 
                 Y428 
                 HQYSDYDyHSSSEKL 
                 SEQ ID NO: 47 
               
               
                   
               
               
                  49 
                 TRAF4 
                 NP_004286.2 
                 Adaptor/scaffold 
                 Y212 
                 EFVFDTIQSHQyQCPR 
                 SEQ ID NO: 48 
               
               
                   
               
               
                  50 
                 CRKL 
                 NP_005198.1 
                 Adaptor/scaffold 
                 Y127 
                 TAEDNLEyVRTLYDF 
                 SEQ ID NO: 49 
               
               
                   
               
               
                  51 
                 ZFYVE9 
                 NP_015563.2 
                 Adaptor/scaffold 
                 Y741 
                 LLyMDRKEARVCVICHSVLMNVAQ 
                 SEQ ID NO: 50 
               
               
                   
                   
                   
                   
                   
                 PR 
               
               
                   
               
               
                  52 
                 TJP1 
                 NP_003248.2 
                 Adaptor/scaffold 
                 Y833 
                 LSYLSAPGSEYSMySTDSR 
                 SEQ ID NO: 51 
               
               
                   
               
               
                  53 
                 TJP2 
                 NP_004808.2 
                 Adaptor/scaffold 
                 Y1178 
                 GyYGQSAR 
                 SEQ ID NO: 52 
               
               
                   
               
               
                  54 
                 LPXN 
                   
                 Adaptor/scaffold; 
                 Y62 
                 PLPAQLVyTTNIQEL 
                 SEQ ID NO: 53 
               
               
                   
                   
                   
                 Cytoskeletal protein 
               
               
                   
               
               
                  55 
                 LPXN 
                   
                 Adaptor/scaffold; 
                 Y72 
                 NIQELNVySEAQEPK 
                 SEQ ID NO: 54 
               
               
                   
                   
                   
                 Cytoskeletal protein 
               
               
                   
               
               
                  56 
                 LPP 
                   
                 Adaptor/scaffold; 
                 Y234 
                 SAQPSPHyMAAPSSG 
                 SEQ ID NO: 55 
               
               
                   
                   
                   
                 Cytoskeletal protein 
               
               
                   
               
               
                  57 
                 LPP 
                 NP_005569.1 
                 Adaptor/scaffold; 
                 Y346 
                 REPGYTPPGAGNQNPPGMyPVTGP 
                 SEQ ID NO: 56 
               
               
                   
                   
                   
                 Cytoskeletal protein 
                   
                 K 
               
               
                   
               
               
                  58 
                 G3BP2 
                 NP_036429.2 
                 Adaptor/scaffold; 
                 Y175 
                 QENANSGyYEAHPV 
                 SEQ ID NO: 57 
               
               
                   
                   
                   
                 RNA binding protein 
               
               
                   
               
               
                  59 
                 G3BP2 
                 NP_036429.2 
                 Adaptor/scaffold; 
                 Y176 
                 QENANSGYyEAHPV 
                 SEQ ID NO: 58 
               
               
                   
                   
                   
                 RNA binding protein 
               
               
                   
               
               
                  60 
                 ADAM18 
                   
                 Adhesion 
                 Y47 
                 VSERKMIyIITIDGQ 
                 SEQ ID NO: 59 
               
               
                   
               
               
                  61 
                 ADAM18 
                 NP_055052.1 
                 Adhesion 
                 Y197 
                 ALyDYMGSEMMAVTQK 
                 SEQ ID NO: 60 
               
               
                   
               
               
                  62 
                 MLLT4 
                 NP_005927.2 
                 Adhesion 
                 Y202 
                 LAAEVyKDMPETSFTRTISNPEVV 
                 SEQ ID NO: 61 
               
               
                   
                   
                   
                   
                   
                 MK 
               
               
                   
               
               
                  63 
                 CSPG3 
                 NP_004377.1 
                 Adhesion 
                 Y264 
                 NPQELYDVYCFARELGGEVFyVGP 
                 SEQ ID NO: 62 
               
               
                   
                   
                   
                   
                   
                 ARR 
               
               
                   
               
               
                  64 
                 DSC1 
                 NP_004939.1 
                 Adhesion 
                 Y34 
                 VyLRVPSHLQAETLVGKVNLEECL 
                 SEQ ID NO: 63 
               
               
                   
                   
                   
                   
                   
                 K 
               
               
                   
               
               
                  65 
                 DSCAM 
                 NP_001380.2 
                 Adhesion 
                 Y468 
                 ISQMITSEGNVVSyLNISSSQVR 
                 SEQ ID NO: 64 
               
               
                   
               
               
                  66 
                 EDIL3 
                 NP_005702.3 
                 Adhesion 
                 Y250 
                 IGSPEYIKSYKIAySNDGKTWAMY 
                 SEQ ID NO: 65 
               
               
                   
                   
                   
                   
                   
                 K 
               
               
                   
               
               
                  67 
                 EDIL3 
                 NP_005702.3 
                 Adhesion 
                 Y260 
                 IGSPEYIKSYKIAYSNDGKTWAMy 
                 SEQ ID NO: 66 
               
               
                   
                   
                   
                   
                   
                 K 
               
               
                   
               
               
                  68 
                 ERBB2IP 
                 NP_001006600.1 
                 Adhesion 
                 Y977 
                 GPTSGPQSAPQIYGPPQyNIQYSS 
                 SEQ ID NO: 67 
               
               
                   
                   
                   
                   
                   
                 SAAVK 
               
               
                   
               
               
                  69 
                 FGL1 
                 NP_004458.3 
                 Adhesion 
                 Y80 
                 RQyADCSEIFNDGYK 
                 SEQ ID NO: 68 
               
               
                   
               
               
                  70 
                 LGALS8 
                 NP_006490.3 
                 Adhesion 
                 Y332 
                 EFKVAVNGVHSLEyKHR 
                 SEQ ID NO: 69 
               
               
                   
               
               
                  71 
                 ICAM2 
                 NP_00864.1 
                 Adhesion 
                 Y260 
                 MGTyGVRAAWRR 
                 SEQ ID NO: 70 
               
               
                   
               
               
                  72 
                 PPFIA1 
                 NP_003617.1 
                 Adhesion 
                 Y546 
                 FPMADGHTDSySTSAVLR 
                 SEQ ID NO: 71 
               
               
                   
               
               
                  73 
                 PECAM1 
                 NP_000433.3 
                 Adhesion 
                 Y663 
                 MSDPNMEANSHyGHNDDVR 
                 SEQ ID NO: 72 
               
               
                   
               
               
                  74 
                 SIGLEC6 
                 NP_001236.3 
                 Adhesion 
                 Y426 
                 SDHPAEAGPISEDEQELHy 
                 SEQ ID NO: 73 
               
               
                   
               
               
                  75 
                 SIGLEC6 
                 NP_001236.3 
                 Adhesion 
                 Y446 
                 VQPQEPKVTDTEySEIK 
                 SEQ ID NO: 74 
               
               
                   
               
               
                  76 
                 SCARF1 
                 NP_003684.2 
                 Adhesion 
                 Y692 
                 TVAEHVEAIEGSVQESSGPVTTIy 
                 SEQ ID NO: 75 
               
               
                   
                   
                   
                   
                   
                 MLAGKPR 
               
               
                   
               
               
                  77 
                 THBS1 
                 NP_003237.2 
                 Adhesion 
                 Y1126 
                 LSHRPKTGFIRVVMyEGK 
                 SEQ ID NO: 76 
               
               
                   
               
               
                  78 
                 URP2 
                 NP_113659.3 
                 Adhesion 
                 Y500 
                 TGSGGPGNHPHGPDASAEGLNPyG 
                 SEQ ID NO: 77 
               
               
                   
                   
                   
                   
                   
                 LVAPR 
               
               
                   
               
               
                  79 
                 ICAM3 
                 NP_002153.1 
                 Adhesion; Immunoglobulin 
                 Y527 
                 EESTyLPLTSMQPTEAMGEEPSRA 
                 SEQ ID NO: 78 
               
               
                   
                   
                   
                 superfamily 
                   
                 E 
               
               
                   
               
               
                  80 
                 DFFA 
                 NP_004392.1 
                 Apoptosis 
                 Y75 
                 DGTIVDDDDyFLCLPSNTKFVALA 
                 SEQ ID NO: 79 
               
               
                   
                   
                   
                   
                   
                 SNE 
               
               
                   
               
               
                  81 
                 PDCD5 
                 NP_004699.1 
                 Apoptosis 
                 Y125 
                 RKVMDSDEDDDy 
                 SEQ ID NO: 80 
               
               
                   
               
               
                  82 
                 HRC 
                 NP_002143.1 
                 Calcium-binding protein 
                 Y209 
                 EEEEEEEEEEEEASTEyGHQAHRH 
                 SEQ ID NO: 81 
               
               
                   
               
               
                  83 
                 CDC45L 
                 NP_003495.1 
                 Cell cycle regulation 
                 Y413 
                 SNLDKLyHGLELAK 
                 SEQ ID NO: 82 
               
               
                   
               
               
                  84 
                 CLASP1 
                 NP_056097.1 
                 Cell cycle regulation 
                 Y697 
                 LLGSGyGGLTGGSSRGPPVTPSSE 
                 SEQ ID NO: 83 
               
               
                   
                   
                   
                   
                   
                 K 
               
               
                   
               
               
                  85 
                 SUGT1 
                 NP_006695.1 
                 Cell cycle regulation 
                 Y47 
                 ALEQKPDDAQyYCQR 
                 SEQ ID NO: 84 
               
               
                   
               
               
                  86 
                 SMC4L1 
                 NP_001002799.1 
                 Cell cycle regulation 
                 Y150 
                 IIDKEGDDyEVIPNSNFYVSR 
                 SEQ ID NO: 85 
               
               
                   
               
               
                  87 
                 TSC2 
                 NP_000539.1 
                 Cell cycle regulation; Tumor 
                 Y1736 
                 SNPTDIyPSKWIARLRHIK 
                 SEQ ID NO: 86 
               
               
                   
                   
                   
                 suppressor; GTPase activating 
               
               
                   
                   
                   
                 protein, misc. 
               
               
                   
               
               
                  88 
                 CD22 
                   
                 Cell surface 
                 Y796 
                 TGDAESSEMQRPPPDCDDTVTySA 
                 SEQ ID NO: 87 
               
               
                   
                   
                   
                   
                   
                 LHKR 
               
               
                   
               
               
                  89 
                 LY9 
                 NP_002339.2 
                 Cell surface 
                 Y626 
                 TPVSQKEESSATIyCSIR 
                 SEQ ID NO: 88 
               
               
                   
               
               
                  90 
                 CD72 
                 NP_001773.1 
                 Cell surface 
                 Y39 
                 LGQDPGADDDGEITyENVQVPAVL 
                 SEQ ID NO: 89 
               
               
                   
                   
                   
                   
                   
                 GVPSSLASSVLGDK 
               
               
                   
               
               
                  91 
                 APLP2 
                 NP_001633.1 
                 Cell surface; DNA binding 
                 Y757 
                 MQNHGYENPTYKyLEQMQI 
                 SEQ ID NO: 90 
               
               
                   
                   
                   
                 protein; Receptor, misc. 
               
               
                   
               
               
                  92 
                 CD84 
                   
                 Cell surface; Immunoglobulin 
                 Y262 
                 AASKKTIyTYIMASR 
                 SEQ ID NO: 91 
               
               
                   
                   
                   
                 superfamily 
               
               
                   
               
               
                  93 
                 CD84 
                   
                 Cell surface; Immunoglobulin 
                 Y279 
                 QPAESRIyDEILQSK 
                 SEQ ID NO: 92 
               
               
                   
                   
                   
                 superfamily 
               
               
                   
               
               
                  94 
                 CD84 
                 NP_003865.1 
                 Cell surface; Immunoglobulin 
                 Y299 
                 VLPSKEEPVNTVySEVQFADKMGK 
                 SEQ ID NO: 93 
               
               
                   
                   
                   
                 superfamily 
               
               
                   
               
               
                  95 
                 CACNB3 
                 NP_000716.2 
                 Channel, calcium 
                 Y429 
                 HLEEDyADAYQDLYQPHR 
                 SEQ ID NO: 94 
               
               
                   
               
               
                  96 
                 CACNB3 
                 NP_000716.2 
                 Channel, calcium 
                 Y433 
                 HLEEDYADAyQDLYQPHR 
                 SEQ ID NO: 95 
               
               
                   
               
               
                  97 
                 CACNB3 
                 NP_000716.2 
                 Channel, calcium 
                 Y437 
                 HLEEDYADAYQDLyQPHR 
                 SEQ ID NO: 96 
               
               
                   
               
               
                  98 
                 MGC15619 
                 NP_115745.2 
                 Channel, cation 
                 Y35 
                 HFTVVGDDyHAWNINYKK 
                 SEQ ID NO: 97 
               
               
                   
               
               
                  99 
                 MGC15619 
                 NP_115745.2 
                 Channel, cation 
                 Y42 
                 FLRHFTVVGDDYHAWNINyK 
                 SEQ ID NO: 98 
               
               
                   
               
               
                 100 
                 GABRA1 
                 NP_000797.2 
                 Channel, chloride 
                 Y53 
                 ILFRLLDGyDNRLRPGLGER 
                 SEQ ID NO: 99 
               
               
                   
               
               
                 101 
                 GABRA1 
                 NP_000797.2 
                 Channel, chloride 
                 Y237 
                 NQYDLLGQTVDSGIVQSSTGEyVV 
                 SEQ ID NO: 100 
               
               
                   
                   
                   
                   
                   
                 MTTHFH 
               
               
                   
               
               
                 102 
                 CFTR 
                 NP_000483.3 
                 Channel, chloride; 
                 Y1307 
                 NLDPyEQWSDQEIWKVADEVGLR 
                 SEQ ID NO: 101 
               
               
                   
                   
                   
                 Transporter, ABC 
               
               
                   
               
               
                 103 
                 P2RX7 
                 NP_002553.2 
                 Channel, ligand-gated; 
                 Y288 
                 TTNVSLyPGYNFRYAK 
                 SEQ ID NO: 102 
               
               
                   
                   
                   
                 Receptor, misc. 
               
               
                   
               
               
                 104 
                 P2RX7 
                 NP_002553.2 
                 Channel, ligand-gated; 
                 Y295 
                 TTNVSLYPGYNFRyAK 
                 SEQ ID NO: 103 
               
               
                   
                   
                   
                 Receptor, misc. 
               
               
                   
               
               
                 105 
                 GJA5 
                 NP_005257.2 
                 Channel, misc. 
                 Y123 
                 EAERAKEVRGSGSyEYPVAEK 
                 SEQ ID NO: 104 
               
               
                   
               
               
                 106 
                 GJA5 
                 NP_005257.2 
                 Channel, misc. 
                 Y125 
                 EAERAKEVRGSGSYEyPVAEK 
                 SEQ ID NO: 105 
               
               
                   
               
               
                 107 
                 CCT2 
                 NP_006422.1 
                 Chaperone 
                 Y297 
                 FINRQLIyNYPEQLF 
                 SEQ ID NO: 106 
               
               
                   
               
               
                 108 
                 DNAJC13 
                 NP_056083.2 
                 Chaperone 
                 Y1024 
                 MLNSNTESPyLIWNNSTR 
                 SEQ ID NO: 107 
               
               
                   
               
               
                 109 
                 FKBP8 
                 NP_036313.3 
                 Chaperone 
                 Y365 
                 STETALyR 
                 SEQ ID NO: 108 
               
               
                   
               
               
                 110 
                 HSPCA 
                 NP_001017963.1 
                 Chaperone 
                 Y319 
                 VILHLKEDQTEyLEER 
                 SEQ ID NO: 109 
               
               
                   
               
               
                 111 
                 HSPCA 
                 NP_001017963.1 
                 Chaperone 
                 Y614 
                 NQKHIyYITGE 
                 SEQ ID NO: 110 
               
               
                   
               
               
                 112 
                 MRCL3 
                 NP_006462.1 
                 Contractile protein 
                 Y155 
                 GNFNyIEFTR 
                 SEQ ID NO: 111 
               
               
                   
               
               
                 113 
                 IL32 
                 NP_001012649.1 
                 Cytokine 
                 Y62 
                 TVAAYyEEQHPE 
                 SEQ ID NO: 112 
               
               
                   
               
               
                 114 
                 ACTN4 
                 NP_004915.2 
                 Cytoskeletal protein 
                 Y234 
                 KDDPVTNLNNAFEVAEKyLDIPK 
                 SEQ ID NO: 113 
               
               
                   
               
               
                 115 
                 ADD1 
                 NP_001110.2 
                 Cytoskeletal protein 
                 Y35 
                 YFDRVDENNPEyLRER 
                 SEQ ID NO: 114 
               
               
                   
               
               
                 116 
                 ADD3 
                 NP_001112.2 
                 Cytoskeletal protein 
                 Y446 
                 WLNSPNTyMK 
                 SEQ ID NO: 115 
               
               
                   
               
               
                 117 
                 BICD2 
                 NP_001003800.1 
                 Cytoskeletal protein 
                 Y425 
                 RQTALDNEKDRDSHEDGDYyEVDI 
                 SEQ ID NO: 116 
               
               
                   
                   
                   
                   
                   
                   
                 NGPE 
               
               
                   
               
               
                 118 
                 KRT6A 
                 NP_005545.1 
                 Cytoskeletal protein 
                 Y278 
                 DVDAAyMNKVELQAK 
                 SEQ ID NO: 117 
               
               
                   
               
               
                 119 
                 CLASP2 
                 NP_055912.1 
                 Cytoskeletal protein 
                 Y1231 
                 SRDyNPYNYSDSISPFNK 
                 SEQ ID NO: 118 
               
               
                   
               
               
                 120 
                 CLASP2 
                 NP_055912.1 
                 Cytoskeletal protein 
                 Y1236 
                 SRDYNPYNySDSISPFNK 
                 SEQ ID NO: 119 
               
               
                   
               
               
                 121 
                 CFL1 
                   
                 Cytoskeletal protein 
                 Y68 
                 GQTVDDPyATFVKML 
                 SEQ ID NO: 120 
               
               
                   
               
               
                 122 
                 CORO1A 
                 NP_009005.1 
                 Cytoskeletal protein 
                 Y180 
                 TLGPEVHPDTIySVDW 
                 SEQ ID NO: 121 
               
               
                   
               
               
                 123 
                 CORO1A 
                 NP_009005.1 
                 Cytoskeletal protein 
                 Y364 
                 KSDLFQEDLyPPTAGPDPALTAEE 
                 SEQ ID NO: 122 
               
               
                   
                   
                   
                   
                   
                 WLGGR 
               
               
                   
               
               
                 124 
                 JUP 
                 NP_002221.1 
                 Cytoskeletal protein 
                 Y660 
                 ISEDKNPDyR 
                 SEQ ID NO: 123 
               
               
                   
               
               
                 125 
                 EMD 
                 NP_000108.1 
                 Cytoskeletal protein 
                 Y90 
                 KEDALLYQSKGyNDDYYEESYFTT 
                 SEQ ID NO: 124 
               
               
                   
                   
                   
                   
                   
                 R 
               
               
                   
               
               
                 126 
                 EMD 
                   
                 Cytoskeletal protein 
                 Y155 
                 KDRERPMyGRDSAYQ 
                 SEQ ID NO: 125 
               
               
                   
               
               
                 127 
                 EMD 
                   
                 Cytoskeletal protein 
                 Y161 
                 MYGRDSAyQSITHYR 
                 SEQ ID NO: 126 
               
               
                   
               
               
                 128 
                 EMD 
                   
                 Cytoskeletal protein 
                 Y181 
                 RSSLDLSyYPTSSST 
                 SEQ ID NO: 127 
               
               
                   
               
               
                 129 
                 ELMO1 
                   
                 Cytoskeletal protein 
                 Y720 
                 EPSNyDFVYDCN 
                 SEQ ID NO: 128 
               
               
                   
               
               
                 130 
                 ELMO1 
                 NP_055615.8 
                 Cytoskeletal protein 
                 Y724 
                 EPSNYDFVyDCN 
                 SEQ ID NO: 129 
               
               
                   
               
               
                 131 
                 EPLIN 
                 NP_057441.1 
                 Cytoskeletal protein 
                 Y190 
                 yNVPLNRLKMMFEKGEPTQTK 
                 SEQ ID NO: 130 
               
               
                   
               
               
                 132 
                 EPLIN 
                 NP_057441.1 
                 Cytoskeletal protein 
                 Y751 
                 NRyYDEDEDEE 
                 SEQ ID NO: 131 
               
               
                   
               
               
                 133 
                 EPB41L2 
                 NP_001422.1 
                 Cytoskeletal protein 
                 Y88 
                 SyTLVVAK 
                 SEQ ID NO: 132 
               
               
                   
               
               
                 134 
                 EPB41L2 
                 NP_001422.1 
                 Cytoskeletal protein 
                 Y773 
                 VTEGTIREEQEyEEEVEEEPRPAA 
                 SEQ ID NO: 133 
               
               
                   
                   
                   
                   
                   
                 K 
               
               
                   
               
               
                 135 
                 KRT2A 
                 NP_000414.2 
                 Cytoskeletal protein 
                 Y463 
                 LNDLEEALQQAKEDLARLLRDyQE 
                 SEQ ID NO: 134 
               
               
                   
                   
                   
                   
                   
                 LMNVK 
               
               
                   
               
               
                 136 
                 LMNB1 
                 NP_005564.1 
                 Cytoskeletal protein 
                 Y482 
                 NTSEQDQPMGGWEMIRKIGDTSVS 
                 SEQ ID NO: 135 
               
               
                   
                   
                   
                   
                   
                 yK 
               
               
                   
               
               
                 137 
                 MAP1A 
                 NP_002364.5 
                 Cytoskeletal protein 
                 Y1388 
                 VVEPKDTAIyQKDE 
                 SEQ ID NO: 136 
               
               
                   
               
               
                 138 
                 MAP1A 
                 NP_002364.5 
                 Cytoskeletal protein 
                 Y1696 
                 GREDVALEQDTyWRELSCER 
                 SEQ ID NO: 137 
               
               
                   
               
               
                 139 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1870 
                 TPGDFSyAYQKPEETTR 
                 SEQ ID NO: 138 
               
               
                   
               
               
                 140 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1872 
                 TPGDFSYAyQKPEETTRSPDEEDY 
                 SEQ ID NO: 139 
               
               
                   
                   
                   
                   
                   
                 DYESYEK 
               
               
                   
               
               
                 141 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1892 
                 SPDEEDYDYESyEK 
                 SEQ ID NO: 140 
               
               
                   
               
               
                 142 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1904 
                 TSDVGGyYYEK 
                 SEQ ID NO: 141 
               
               
                   
               
               
                 143 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1905 
                 TSDVGGYyYEK 
                 SEQ ID NO: 142 
               
               
                   
               
               
                 144 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1921 
                 SPSDSGySYETIGK 
                 SEQ ID NO: 143 
               
               
                   
               
               
                 145 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1955 
                 TPEEGGySYDISEK 
                 SEQ ID NO: 144 
               
               
                   
               
               
                 146 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1957 
                 TPEEGGYSyDISEK 
                 SEQ ID NO: 145 
               
               
                   
               
               
                 147 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1972 
                 TTSPPEVSGySYEK 
                 SEQ ID NO: 146 
               
               
                   
               
               
                 148 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1974 
                 TTSPPEVSGYSyEK 
                 SEQ ID NO: 147 
               
               
                   
               
               
                 149 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y1991 
                 LLDDISNGyDDSEDGGHTLGDPSY 
                 SEQ ID NO: 148 
               
               
                   
                   
                   
                   
                   
                 SYETTEK 
               
               
                   
               
               
                 150 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y2006 
                 LLDDISNGYDDSEDGGHTLGDPSy 
                 SEQ ID NO: 149 
               
               
                   
                   
                   
                   
                   
                 SYETTEK 
               
               
                   
               
               
                 151 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y2008 
                 LLDDISNGYDDSEDGGHTLGDPSY 
                 SEQ ID NO: 150 
               
               
                   
                   
                   
                   
                   
                 SyETTEK 
               
               
                   
               
               
                 152 
                 MAP1B 
                 NP_005900.1 
                 Cytoskeletal protein 
                 Y2025 
                 ITSFPESEGYSyETSTK 
                 SEQ ID NO: 151 
               
               
                   
               
               
                 153 
                 MAP2 
                   
                 Cytoskeletal protein 
                 Y1685 
                 yQPKGGQVR 
                 SEQ ID NO: 152 
               
               
                   
               
               
                 154 
                 MAP4 
                 NP_002366.2 
                 Cytoskeletal protein 
                 Y1001 
                 VSySHIQSK 
                 SEQ ID NO: 153 
               
               
                   
               
               
                 155 
                 NEB 
                 NP_004534.1 
                 Cytoskeletal protein 
                 Y4112 
                 AYELQSDNVyKADLEWLR 
                 SEQ ID NO: 154 
               
               
                   
               
               
                 156 
                 PLEC1 
                 NP_000436.2 
                 Cytoskeletal protein 
                 Y4283 
                 SRSSSVGSSSSyPISPAVSR 
                 SEQ ID NO: 155 
               
               
                   
               
               
                 157 
                 RSN 
                 NP_002947.1 
                 Cytoskeletal protein 
                 Y108 
                 NDGSVAGVRyFQCEPLK 
                 SEQ ID NO: 156 
               
               
                   
               
               
                 158 
                 TAGLN2 
                 NP_003555.1 
                 Cytoskeletal protein 
                 Y8 
                 GPAyGLSR 
                 SEQ ID NO: 157 
               
               
                   
               
               
                 159 
                 SPTBN1 
                 NP_842565.1 
                 Cytoskeletal protein 
                 Y17 
                 TSSISGPLSPAyTGQVPYNYNQLE 
                 SEQ ID NO: 158 
               
               
                   
                   
                   
                   
                   
                 GR 
               
               
                   
               
               
                 160 
                 TLN1 
                 NP_006280.2 
                 Cytoskeletal protein 
                 Y127 
                 IGITNHDEySLVR 
                 SEQ ID NO: 159 
               
               
                   
               
               
                 161 
                 HRIHFB2 
                 NP_008963.3 
                 Cytoskeletal protein 
                 Y553 
                 FTSGKYQDVyVELSHIK 
                 SEQ ID NO: 160 
               
               
                   
                 122 
               
               
                   
               
               
                 162 
                 TUBA1 
                   
                 Cytoskeletal protein 
                 Y210 
                 DNEAIyDICRRNLDIERPT 
                 SEQ ID NO: 161 
               
               
                   
               
               
                 163 
                 TUBA1 
                   
                 Cytoskeletal protein 
                 Y224 
                 NLDIERPTyTNLNR 
                 SEQ ID NO: 162 
               
               
                   
               
               
                 164 
                 TUBA1 
                 NP_005991.1 
                 Cytoskeletal protein 
                 Y282 
                 AyHEQLSVAEITNACFEPANQMVK 
                 SEQ ID NO: 163 
               
               
                   
               
               
                 165 
                 TUBA1 
                 NP_005991.1 
                 Cytoskeletal protein 
                 Y399 
                 LDHKFDLMyAKR 
                 SEQ ID NO: 164 
               
               
                   
               
               
                 166 
                 TUBA3 
                 NP_006000.2 
                 Cytoskeletal protein 
                 Y451 
                 EDMAALEKDYEEVGVGSVEGEGEE 
                 SEQ ID NO: 165 
               
               
                   
                   
                   
                   
                   
                 EGEEy 
               
               
                   
               
               
                 167 
                 TUBB2 
                 NP_001060.1 
                 Cytoskeletal protein 
                 Y106 
                 GHyTEGAELVDSVLDVVRK 
                 SEQ ID NO: 166 
               
               
                   
               
               
                 168 
                 TUBB2 
                 NP_001060.1 
                 Cytoskeletal protein 
                 Y222 
                 LTTPTyGDLNHLVSATMSGVTTCL 
                 SEQ ID NO: 167 
               
               
                   
                   
                   
                   
                   
                 R 
               
               
                   
               
               
                 169 
                 TUBB 
                 NP_821133.1 
                 Cytoskeletal protein 
                 Y51 
                 ISVYyNEATGGK 
                 SEQ ID NO: 168 
               
               
                   
               
               
                 170 
                 VIM 
                 NP_003371.2 
                 Cytoskeletal protein 
                 Y276 
                 DVRQQyESVAAK 
                 SEQ ID NO: 169 
               
               
                   
               
               
                 171 
                 RAD51 
                 NP_002866.2 
                 DNA repair 
                 Y232 
                 YALLIVDSATALYRTDySGRGELS 
                 SEQ ID NO: 170 
               
               
                   
                   
                   
                   
                   
                 ARQMHLAR 
               
               
                   
               
               
                 172 
                 XRCC1 
                 NP_006288.1 
                 DNA repair 
                 Y576 
                 RKLIRYVTAFNGELEDyMSDR 
                 SEQ ID NO: 171 
               
               
                   
               
               
                 173 
                 RFC2 
                 NP_002905.2 
                 DNA replication 
                 Y277 
                 TFQMAEyLKLEFIKEIGYTHMK 
                 SEQ ID NO: 172 
               
               
                   
               
               
                 174 
                 GIMAP1 
                 NP_570115.1 
                 G protein regulator, misc. 
                 Y14 
                 MATDEENVyGLEENAQSR 
                 SEQ ID NO: 173 
               
               
                   
               
               
                 175 
                 ARHGDIA 
                 NP_004300.1 
                 G protein regulator, misc. 
                 Y27 
                 HSVNyKPPAQKSIQE 
                 SEQ ID NO: 174 
               
               
                   
               
               
                 176 
                 ARHGDIB 
                 NP_001166.3 
                 G protein regulator, misc. 
                 Y48 
                 SLKELQEMDKDDESLIKyK 
                 SEQ ID NO: 175 
               
               
                   
               
               
                 177 
                 SIPA1L3 
                 NP_055888.1 
                 G protein regulator, misc. 
                 Y1068 
                 RPVSFPETPyTVSPAGADR 
                 SEQ ID NO: 176 
               
               
                   
               
               
                 178 
                 GNA12 
                 NP_002061.1 
                 G protein, heterotrimeric 
                 Y61 
                 IIHEDGySEEECR 
                 SEQ ID NO: 177 
               
               
                   
               
               
                 179 
                 GNA15 
                 NP_002059.1 
                 G protein, heterotrimeric 
                 Y83 
                 QMRIIHGAGYSEEERKGFRPLVyQ 
                 SEQ ID NO: 178 
               
               
                   
                   
                   
                   
                   
                 NIFVSMR 
               
               
                   
               
               
                 180 
                 GNA13 
                 NP_006487.1 
                 G protein, heterotrimeric 
                 Y354 
                 NNLKECGLy 
                 SEQ ID NO: 179 
               
               
                   
               
               
                 181 
                 ARFGAP1 
                 NP_060679.1 
                 GTPase activating protein, 
                 Y208 
                 GNTPPPQKKEDDFLNNAMSSLySG 
                 SEQ ID NO: 180 
               
               
                   
                   
                   
                 ARF 
                   
                 W 
               
               
                   
               
               
                 182 
                 ARFGAP3 
                   
                 GTPase activating protein, 
                 Y378 
                 WDDSSDSyWKKETSK 
                 SEQ ID NO: 181 
               
               
                   
                   
                   
                 ARF 
               
               
                   
               
               
                 183 
                 CENTB1 
                 NP_055531.1 
                 GTPase activating protein, 
                 Y712 
                 EAEAAQGQAGDETyLDIFR 
                 SEQ ID NO: 182 
               
               
                   
                   
                   
                 ARF 
               
               
                   
               
               
                 184 
                 CENTD3 
                 NP_071926.4 
                 GTPase activating protein, 
                 Y303 
                 LTPLLSGWLDKLSPQGNyVFQR 
                 SEQ ID NO: 183 
               
               
                   
                   
                   
                 ARF; GTPase activating 
               
               
                   
                   
                   
                 protein, Rac/Rho 
               
               
                   
               
               
                 185 
                 CENTD3 
                 NP_071926.4 
                 GTPase activating protein, 
                 Y139 
                 SLMyFGSDKDPFPK 
                 SEQ ID NO: 184 
               
               
                   
                   
                   
                 ARF; GTPase activating 
               
               
                   
                   
                   
                 protein, Rac/Rho 
               
               
                   
               
               
                 186 
                 CENTD3 
                 NP_071926.4 
                 GTPase activating protein, 
                 Y489 
                 QSWAAALQEAVTETLSDyEVAEK 
                 SEQ ID NO: 185 
               
               
                   
                   
                   
                 ARF; GTPase activating 
               
               
                   
                   
                   
                 protein, Rac/Rho 
               
               
                   
               
               
                 187 
                 CENTD3 
                 NP_071926.4 
                 GTPase activating protein, 
                 Y684 
                 ATYSGFLyCSPVSNK 
                 SEQ ID NO: 186 
               
               
                   
                   
                   
                 ARF; GTPase activating 
               
               
                   
                   
                   
                 protein, Rac/Rho 
               
               
                   
               
               
                 188 
                 CENTD3 
                 NP_071926.4 
                 GTPase activating protein, 
                 Y882 
                 TLyGQGEGR 
                 SEQ ID NO: 187 
               
               
                   
                   
                   
                 ARF; GTPase activating 
               
               
                   
                   
                   
                 protein, Rac/Rho 
               
               
                   
               
               
                 189 
                 SIPA1L1 
                 NP_056371.1 
                 GTPase activating protein, 
                 Y1166 
                 TGSVGGTyRQKSMPE 
                 SEQ ID NO: 188 
               
               
                   
                   
                   
                 misc. 
               
               
                   
               
               
                 190 
                 ARHGAP4 
                 NP_001657.2 
                 GTPase activating protein, 
                 Y483 
                 GDKEEQEVSWTQyTQRK 
                 SEQ ID NO: 189 
               
               
                   
                   
                   
                 Rac/Rho 
               
               
                   
               
               
                 191 
                 RGS10 
                 NP_001005339.1 
                 GTPase activating protein, 
                 Y94 
                 EIyMTFLSSKASSQVNVEGQSR 
                 SEQ ID NO: 190 
               
               
                   
                   
                   
                 RGS 
               
               
                   
               
               
                 192 
                 RGS14 
                 NP_006471.2 
                 GTPase activating protein, 
                 Y122 
                 NIyQEFLSSQALSPVNIDR 
                 SEQ ID NO: 191 
               
               
                   
                   
                   
                 RGS 
               
               
                   
               
               
                 193 
                 ARL1 
                 NP_001168.1 
                 Guanine nucleotide exchange 
                 Y58 
                 NVETVTyKNLKFQVW 
                 SEQ ID NO: 192 
               
               
                   
                   
                   
                 factor, ARF 
               
               
                   
               
               
                 194 
                 CENTD2 
                 NP_056057.1 
                 Guanine nucleotide exchange 
                 Y191 
                 ARLSSAyLLGVPGSEQPDRAGSLE 
                 SEQ ID NO: 193 
               
               
                   
                   
                   
                 factor, ARF 
                   
                 LR 
               
               
                   
               
               
                 195 
                 WBSCR16 
                   
                 Guanine nucleotide exchange 
                 Y216 
                 KVVENEIySESHRVH 
                 SEQ ID NO: 194 
               
               
                   
                   
                   
                 factor, misc. 
               
               
                   
               
               
                 196 
                 ARHGEF6 
                 NP_004831.1 
                 Guanine nucleotide exchange 
                 Y644 
                 KPSEEEyVIRK 
                 SEQ ID NO: 195 
               
               
                   
                   
                   
                 factor, Rac/Rho 
               
               
                   
               
               
                 197 
                 RAC2 
                 NP_002863.1 
                 Guanine nucleotide exchange 
                 Y139 
                 EKKLAPITyPQGLALAKEIDSVK 
                 SEQ ID NO: 196 
               
               
                   
                   
                   
                 factor, Rac/Rho 
               
               
                   
               
               
                 198 
                 ARHGEF2 
                 NP_004714.2 
                 Guanine nucleotide exchange 
                 Y866 
                 SLPAGDALyLSFNPPQPSR 
                 SEQ ID NO: 197 
               
               
                   
                   
                   
                 factor, Rac/Rho 
               
               
                   
               
               
                 199 
                 VAV1 
                   
                 Guanine nucleotide exchange 
                 Y826 
                 GWWRGEIyGRVGWFP 
                 SEQ ID NO: 198 
               
               
                   
                   
                   
                 factor, Rac/Rho 
               
               
                   
               
               
                 200 
                 RAPGEF1 
                   
                 Guanine nucleotide exchange 
                 Y341 
                 RLSGGSHSyGGESPRLSPCSSIDK 
                 SEQ ID NO: 199 
               
               
                   
                   
                   
                 factor, Ras 
                   
                 LSK 
               
               
                   
               
               
                 201 
                 DDX46 
                 NP_055644.2 
                 Helicase 
                 Y730 
                 yAGDIIKALELSGTAVPPDLEK 
                 SEQ ID NO: 200 
               
               
                   
               
               
                 202 
                 DDX3X 
                 NP_001347.3 
                 Helicase 
                 Y580 
                 QEVPSWLENMAYEHHyK 
                 SEQ ID NO: 201 
               
               
                   
               
               
                 203 
                 DDX20 
                 NP_009135.3 
                 Helicase 
                 Y756 
                 LQTEAQEDDWyDCHR 
                 SEQ ID NO: 202 
               
               
                   
               
               
                 204 
                 BAT1 
                 NP_004631.1 
                 Helicase 
                 Y39 
                 GSyVSIHSSGFR 
                 SEQ ID NO: 203 
               
               
                   
               
               
                 205 
                 WRN 
                 NP_000544.2 
                 Helicase 
                 Y212 
                 LyAATDAYAGFIIYR 
                 SEQ ID NO: 204 
               
               
                   
               
               
                 206 
                 CD7 
                 NP_006128.1 
                 Immunoglobulin superfamily 
                 Y239 
                 CNTLSSPNQyQ 
                 SEQ ID NO: 205 
               
               
                   
               
               
                 207 
                 SLAMF9 
                 NP_254273.1 
                 Immunoglobulin superfamily 
                 Y125 
                 TSQISTMQQyNLCVYR 
                 SEQ ID NO: 206 
               
               
                   
               
               
                 208 
                 ANP32A 
                 NP_006296.1 
                 Inhibitor protein 
                 Y148 
                 LLPQLTYLDGyDR 
                 SEQ ID NO: 207 
               
               
                   
               
               
                 209 
                 SERPINC1 
                 NP_000479.1 
                 Inhibitor protein 
                 Y163 
                 TSDQIHFFFAKLNCRLyR 
                 SEQ ID NO: 208 
               
               
                   
               
               
                 210 
                 AK2 
                   
                 Kinase (non-protein) 
                 Y200 
                 IRLQAYHTQTTPLIEyYRK 
                 SEQ ID NO: 209 
               
               
                   
               
               
                 211 
                 CMPK 
                 NP_057392.1 
                 Kinase (non-protein) 
                 Y187 
                 RIQTyLQSTKPIIDLYEEMGKVKK 
                 SEQ ID NO: 210 
               
               
                   
                   
                   
                   
                   
                 IDASK 
               
               
                   
               
               
                 212 
                 CMPK 
                 NP_057392.1 
                 Kinase (non-protein) 
                 Y198 
                 RIQTYLQSTKPIIDLyEEMGKVKK 
                 SEQ ID NO: 212 
               
               
                   
                   
                   
                   
                   
                 IDASK 
               
               
                   
               
               
                 213 
                 HK1 
                 NP_000179.1 
                 Kinase (non-protein) 
                 Y749 
                 MISGMyLGEIVR 
                 SEQ ID NO: 212 
               
               
                   
               
               
                 214 
                 PRPS1 
                 NP_002755.1 
                 Kinase (non-protein) 
                 Y146 
                 QGFFDIPVDNLyAEPA 
                 SEQ ID NO: 213 
               
               
                   
               
               
                 215 
                 TRIM24 
                 NP_003843.3 
                 KINASE(atypical); Protein 
                 Y506 
                 yPPNQNIPRQAIKPNPLQMAFLAQ 
                 SEQ ID NO: 214 
               
               
                   
                   
                   
                 kinase 
                   
                 QAIK 
               
               
                   
               
               
                 216 
                 TRIM28 
                 NP_005753.1 
                 KINASE(atypical); Protein 
                 Y242 
                 DCQLNAHKDHQyQFLEDAVR 
                 SEQ ID NO: 215 
               
               
                   
                   
                   
                 kinase 
               
               
                   
               
               
                 217 
                 TRIM28 
                 NP_005753.1 
                 KINASE(atypical); Protein 
                 Y369 
                 LIyFQLHR 
                 SEQ ID NO: 216 
               
               
                   
                   
                   
                 kinase 
               
               
                   
               
               
                 218 
                 TRIM33 
                 NP_056990.3 
                 KINASE(atypical); Protein 
                 Y1018 
                 HSQHyQIPDDFVADVRLIFK 
                 SEQ ID NO: 217 
               
               
                   
                   
                   
                 kinase 
               
               
                   
               
               
                 219 
                 PTK9 
                 NP_002813.2 
                 KINASE(dual); Protein kinase, 
                 Y135 
                 QKMLyAATRATLKKEFGGGHIK 
                 SEQ ID NO: 218 
               
               
                   
                   
                   
                 dual-specificity 
               
               
                   
               
               
                 220 
                 DYRK1A 
                 NP_001387.2 
                 KINASE(dual); Protein kinase, 
                 Y177 
                 YEIDSLIGKGSFGQVVKAyDR 
                 SEQ ID NO: 219 
               
               
                   
                   
                   
                 dual-specificity 
               
               
                   
               
               
                 221 
                 NPR2 
                 NP_003986.2 
                 KINASE(dual); Protein kinase, 
                 Y725 
                 SGPFyLEGLDLSPKEIVQK 
                 SEQ ID NO: 220 
               
               
                   
                   
                   
                 dual-specificity 
               
               
                   
               
               
                 222 
                 TTK 
                 NP_003309.2 
                 KINASE(dual); Protein kinase, 
                 Y811 
                 GTTEEMKyVLGQLVGLNSPNSILK 
                 SEQ ID NO: 221 
               
               
                   
                   
                   
                 dual-specificity 
               
               
                   
               
               
                 223 
                 PKIA 
                   
                 KINASE(regulator); Protein 
                 Y8 
                 MTDVETTyADFIASGRTGR 
                 SEQ ID NO: 222 
               
               
                   
                   
                   
                 kinase, regulatory subunit 
               
               
                   
               
               
                 224 
                 PRKAR1B 
                 NP_002726.1 
                 KINASE(regulator); Protein 
                 Y312 
                 SPNEEyVEVGR 
                 SEQ ID NO: 223 
               
               
                   
                   
                   
                 kinase, regulatory subunit 
               
               
                   
               
               
                 225 
                 BCR 
                 NP_004318.3 
                 KINASE(S/T); GTPase 
                 Y58 
                 MIyLQTLLAK 
                 SEQ ID NO: 224 
               
               
                   
                   
                   
                 activating protein, Rac/Rho; 
               
               
                   
                   
                   
                 Protein kinase, Ser/Thr 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 226 
                 BCR 
                 NP_004318.3 
                 KINASE(S/T); GTPase 
                 Y231 
                 SQHGAGSSVGDASRPPyR 
                 SEQ ID NO: 225 
               
               
                   
                   
                   
                 activating protein, Rac/Rho; 
               
               
                   
                   
                   
                 Protein kinase, Ser/Thr 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 227 
                 BCR 
                 NP_004318.3 
                 KINASE(S/T); GTPase 
                 Y554 
                 VPELyEIHKEFYDGLFPR 
                 SEQ ID NO: 226 
               
               
                   
                   
                   
                 activating protein, Rac/Rho; 
               
               
                   
                   
                   
                 Protein kinase, Ser/Thr 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 228 
                 BCR 
                 NP_004318.3 
                 KINASE(S/T); GTPase 
                 Y561 
                 VPELYEIHKEFyDGLFPR 
                 SEQ ID NO: 227 
               
               
                   
                   
                   
                 activating protein, Rac/Rho; 
               
               
                   
                   
                   
                 Protein kinase, Ser/Thr 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 229 
                 BCR 
                 NP_004318.3 
                 KINASE(S/T); GTPase 
                 Y852 
                 SYTFLISSDyER 
                 SEQ ID NO: 228 
               
               
                   
                   
                   
                 activating protein, Rac/Rho; 
               
               
                   
                   
                   
                 Protein kinase, Ser/Thr 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 230 
                 ATM 
                 NP_000042.3 
                 KINASE(S/T); Kinase, lipid; 
                 Y2129 
                 EVEGTSYHESLyNALQSLRDREFS 
                 SEQ ID NO: 229 
               
               
                   
                   
                   
                 Protein kinase, Ser/Thr 
                   
                 TFYESLKYAR 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 231 
                 RIOK2 
                 NP_060813.1 
                 KINASE(S/T); Protein kinase 
                 Y445 
                 VQGGVPAGSDEyEDECPHLIALSS 
                 SEQ ID NO: 230 
               
               
                   
                   
                   
                   
                   
                 LNR 
               
               
                   
               
               
                 232 
                 RIOK2 
                 NP_060813.1 
                 KINASE(S/T); Protein kinase 
                 Y477 
                 EFRPFRDEENVGAMNQyR 
                 SEQ ID NO: 231 
               
               
                   
               
               
                 233 
                 RIPK4 
                 NP_065690.2 
                 KINASE(S/T); Protein kinase, 
                 Y576 
                 GVDVSLQGKDAWLPLHyAAWQGHL 
                 SEQ ID NO: 232 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 PIVKLLAK 
               
               
                   
               
               
                 234 
                 STK6 
                 NP_003591.2 
                 KINASE(S/T); Protein kinase, 
                 Y236 
                 LSKFDEQRTATyITELANALSYCH 
                 SEQ ID NO: 233 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 SK 
               
               
                   
               
               
                 235 
                 STK6 
                 NP_003591.2 
                 KINASE(S/T); Protein kinase, 
                 Y246 
                 LSKFDEQRTATITELANALSyCHS 
                 SEQ ID NO: 234 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 K 
               
               
                   
               
               
                 236 
                 CSNK1A1 
                 NP_001883.4 
                 KINASE(S/T); Protein kinase, 
                 Y294 
                 TLNHQYDyTFDWTMLK 
                 SEQ ID NO: 235 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 237 
                 CDC2L5 
                 NP_003709.2 
                 KINASE(S/T); Protein kinase, 
                 Y300 
                 EPPKAyREDK 
                 SEQ ID NO: 236 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 238 
                 CDKL2 
                 NP_003939.1 
                 KINASE(S/T); Protein kinase, 
                 Y15 
                 YENLGLVGEGSyGMVMKCR 
                 SEQ ID NO: 237 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 239 
                 GRK4 
                 NP_001004056.1 
                 KINASE(S/T); Protein kinase, 
                 Y222 
                 FVVSLAYAyETK 
                 SEQ ID NO: 238 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 240 
                 LATS1 
                 NP_004681.1 
                 KINASE(S/T); Protein kinase, 
                 Y779 
                 DNLyFVMDYIPGGDMMSLLIR 
                 SEQ ID NO: 239 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 241 
                 MARK2 
                 NP_004945.3 
                 KINASE(S/T); Protein kinase, 
                 Y389 
                 FSDQAGPAIPTSNSySKK 
                 SEQ ID NO: 240 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 241 
                 MARK2 
                 NP_004945.3 
                 KINASE(S/T); Protein kinase, 
                 Y389 
                 FSDQAGPAIPTSNSySKK 
                 SEQ ID NO: 240 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                  242 
                 MARK2 
                 NP_004945.3 
                 KINASE(S/T); Protein kinase, 
                 Y525 
                 DQQNLPyGVTPASPSGHSQGR 
                 SEQ ID NO: 241 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 243 
                 MARK3 
                 NP_002367.4 
                 KINASE(S/T); Protein kinase, 
                 Y432 
                 RYSDHAGPAIPSVVAyPK 
                 SEQ ID NO: 242 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 244 
                 MELK 
                 NP_055606.1 
                 KINASE(S/T); Protein kinase, 
                 Y438 
                 SAVKNEEyFMFPEPK 
                 SEQ ID NO: 243 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 245 
                 MAP3K3 
                 NP_002392.2 
                 KINASE(S/T); Protein kinase, 
                 Y155 
                 ASQSAGDINTIyQPPEPR 
                 SEQ ID NO: 244 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 246 
                 MINK1 
                 NP_722549.2 
                 KINASE(S/T); Protein kinase, 
                 Y706 
                 SNSAWQIyLQR 
                 SEQ ID NO: 245 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 247 
                 MINK1 
                 NP_722549.2 
                 KINASE(S/T); Protein kinase, 
                 Y900 
                 GQSPPSKDGSGDyQSR 
                 SEQ ID NO: 246 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 248 
                 MYLK 
                 NP_444253.2 
                 KINASE(S/T); Protein kinase, 
                 Y464 
                 QEGSIEVyEDAGSHYLCLLK 
                 SEQ ID NO: 247 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 249 
                 MYLK 
                 NP_444253.2 
                 KINASE(S/T); Protein kinase, 
                 Y471 
                 QEGSIEVYEDAGSHyLCLLK 
                 SEQ ID NO: 248 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 250 
                 MAPK14 
                 NP_001306.1 
                 KINASE(S/T); Protein kinase, 
                 Y24 
                 yQNLSPVGSGAYGSVCAAFDTKTG 
                 SEQ ID NO: 249 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 LR 
               
               
                   
               
               
                 251 
                 ALS2CR7 
                 NP_631897.1 
                 KINASE(S/T); Protein kinase, 
                 Y63 
                 LGEGSyATVYKGISRINGQLVALK 
                 SEQ ID NO: 250 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 252 
                 PRKCZ 
                 NP_002735.3 
                 KINASE(S/T); Protein kinase, 
                 Y356 
                 FyAAEICIALNFLHER 
                 SEQ ID NO: 251 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 253 
                 MARK3 
                   
                 KINASE(S/T); Protein kinase, 
                 Y402 
                 ySDHAGPGIPSVVAYPKRSQTSTA 
                 SEQ ID NO: 252 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 DSDLK 
               
               
                   
               
               
                 254 
                 STK31 
                 NP_113602.2 
                 KINASE(S/T); Protein kinase, 
                 Y992 
                 YTLyKKEEE 
                 SEQ ID NO: 253 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 255 
                 DKFZp76 
                 XP_291277.2 
                 KINASE(S/T); Protein kinase, 
                 Y132 
                 QEDAPVVyLGSFR 
                 SEQ ID NO: 254 
               
               
                   
                 1P0423 
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 256 
                 STK39 
                 NP_037365.1 
                 KINASE(S/T); Protein kinase, 
                 Y446 
                 QIQSLSVHDSQGPPNANEDyRE 
                 SEQ ID NO: 255 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 257 
                 TSSK1 
                 NP_114417.1 
                 KINASE(S/T); Protein kinase, 
                 Y12 
                 RGyLLGINLGEGSYAKVK 
                 SEQ ID NO: 256 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 258 
                 TTN 
                 NP_003310.3 
                 KINASE(S/T); Protein kinase, 
                 Y1845 
                 SKRFRVRyDGIHYLDIVDCKSYDT 
                 SEQ ID NO: 257 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 GEVK 
               
               
                   
               
               
                 259 
                 TTN 
                 NP_003310.3 
                 KINASE(S/T); Protein kinase, 
                 Y1850 
                 SKRFRVRYDGIHyLDIVDCKSYDT 
                 SEQ ID NO: 258 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 GEVK 
               
               
                   
               
               
                 260 
                 TTN 
                 NP_003310.3 
                 KINASE(S/T); Protein kinase, 
                 Y1859 
                 SKRFRVRYDGIHYLDIVDCKSyDT 
                 SEQ ID NO: 259 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 GEVK 
               
               
                   
               
               
                 261 
                 TTN 
                 NP_003310.3 
                 KINASE(S/T); Protein kinase, 
                 Y8052 
                 DLIONGEyFFR 
                 SEQ ID NO: 260 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 262 
                 KALRN 
                 NP_003938.1 
                 KINASE(S/T); Protein kinase, 
                 Y1351 
                 yEQLPEDVGHCFVTWADKFQMYVT 
                 SEQ ID NO: 261 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 YCKNK 
               
               
                   
               
               
                 263 
                 KALRN 
                 NP_003938.1 
                 KINASE(S/T); Protein kinase, 
                 Y1372 
                 YEQLPEDVGHVFVTWADKFQMyVT 
                 SEQ ID NO: 262 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 YCKNK 
               
               
                   
               
               
                 264 
                 KALRN 
                 NP_003938.1 
                 KINASE(S/T); Protein kinase, 
                 Y1375 
                 YEQLPEDVGHCFVTWADKFQMYVT 
                 SEQ ID NO: 263 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
                   
                 yCKNK 
               
               
                   
               
               
                 265 
                 AKT3 
                 NP_005456.1 
                 KINASE(S/T); Protein kinase, 
                 Y251 
                 TRFyGAEIVSALDYLHSGKIVYR 
                 SEQ ID NO: 264 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 266 
                 AKT3 
                 NP_005456.1 
                 KINASE(S/T); Protein kinase, 
                 Y269 
                 TRFYGAEIVSALDYLHSGKIVyR 
                 SEQ ID NO: 265 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 267 
                 WNK1 
                 NP_061852.1 
                 KINASE(S/T); Protein kinase, 
                 Y2276 
                 GHMNyEGPGMAR 
                 SEQ ID NO: 266 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
               
               
                 268 
                 MAPK3 
                 NP_002737.2 
                 KINASE(S/T); Protein kinase, 
                 Y210 
                 IADPEHDHTGFLTEYVATRWyR 
                 SEQ ID NO: 267 
               
               
                   
                   
                   
                 Ser/Thr (non-receptor) 
               
               
                   
                   
                   
                 Transcription factor 
               
               
                   
               
               
                 269 
                 ACVR2B 
                 NP_001097.1 
                 KINASE(S/T); Receptor 
                 Y85 
                 RLHCYASWANSSGTIELVKKGCWL 
                 SEQ ID NO: 268 
               
               
                   
                   
                   
                 Ser/Thr kinase 
                   
                 DDFNCyDR 
               
               
                   
               
               
                 270 
                 BMPR1A 
                 NP_004320.2 
                 KINASE(S/T); Receptor 
                 Y407 
                 RyMAPEVLDESLNK 
                 SEQ ID NO: 269 
               
               
                   
                   
                   
                 Ser/Thr kinase 
               
               
                   
               
               
                 271 
                 BLK 
                 NP_001706.2 
                 KINASE(Y); Protein kinase, 
                 Y187 
                 CLDEGGyYISPR 
                 SEQ ID NO: 270 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 272 
                 BTK 
                 NP_000052.1 
                 KINASE(Y); Protein kinase, 
                 Y344 
                 HYVVCSTPQSQyYLAEK 
                 SEQ ID NO: 271 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 273 
                 LCK 
                 NP_005347.2 
                 KINASE(Y); Protein kinase, 
                 Y470 
                 MVRPDNCPEELyQLMR 
                 SEQ ID NO: 272 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 274 
                 SYK 
                 NP_003168.2 
                 KINASE(Y); Protein kinase, 
                 Y296 
                 IKSySFPKPGHR 
                 SEQ ID NO: 273 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 275 
                 SYK 
                 NP_003168.2 
                 KINASE(Y); Protein kinase, 
                 Y630 
                 LRNYyYDVVN 
                 SEQ ID NO: 274 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 276 
                 SYK 
                 NP_003168.2 
                 KINASE(Y); Protein kinase, 
                 Y631 
                 LRNYYyDVVN 
                 SEQ ID NO: 275 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 277 
                 TEC 
                 NP_003206.1 
                 KINASE(Y); Protein kinase, 
                 Y519 
                 RYFLDDQyTSSSGAK 
                 SEQ ID NO: 276 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 278 
                 TYK2 
                 NP_003322.2 
                 KINASE(Y); Protein kinase, 
                 Y433 
                 LTADSSHyLCHEVAPPR 
                 SEQ ID NO: 277 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 279 
                 TYK2 
                 NP_003322.2 
                 KINASE(Y); Protein kinase, 
                 Y914 
                 VSLyCYDPTNDGTGEMVAVK 
                 SEQ ID NO: 278 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 280 
                 ABL1 
                 NP_005148.2 
                 KINASE(Y); Protein kinase, 
                 Y128 
                 HSWyHGPVSR 
                 SEQ ID NO: 279 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 281 
                 FES 
                   
                 KINASE(Y); Protein kinase, 
                 Y713 
                 EEADGVyAASGGLR 
                 SEQ ID NO: 280 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 282 
                 LYN 
                 NP_002341.1 
                 KINASE(Y); Protein kinase, 
                 Y266 
                 LGAGQFGEVWMGYyNNSTK 
                 SEQ ID NO: 281 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 283 
                 ZAP70 
                 NP_001070.2 
                 KINASE(Y); Protein kinase, 
                 Y69 
                 QLNGTyAIAGGK 
                 SEQ ID NO: 282 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 284 
                 ZAP70 
                 NP_001070.2 
                 KINASE(Y); Protein kinase, 
                 Y164 
                 MPWyHSSLTR 
                 SEQ ID NO: 283 
               
               
                   
                   
                   
                 tyrosine (non-receptor) 
               
               
                   
               
               
                 285 
                 EPHA7 
                 NP_004431.1 
                 KINASE(Y); Receptor tyrosine 
                 Y511 
                 STSASINNLKPGTVyVFQIR 
                 SEQ ID NO: 284 
               
               
                   
                   
                   
                 kinase, 
               
               
                   
               
               
                 286 
                 FLT3 
                 NP_004110.1 
                 KINASE(Y); Receptor tyrosine 
                 Y630 
                 VMNATAyGISK 
                 SEQ ID NO: 285 
               
               
                   
                   
                   
                 kinase, 
               
               
                   
               
               
                 287 
                 FLT3 
                 NP_004110.1 
                 KINASE(Y); Receptor tyrosine 
                 Y726 
                 TWTEIFKEHNFSFyPTFQSHPNSS 
                 SEQ ID NO: 286 
               
               
                   
                   
                   
                 kinase, 
                   
                 MPGSR 
               
               
                   
               
               
                 288 
                 FLT3 
                 NP_004110.1 
                 KINASE(Y); Receptor tyrosine 
                 Y768 
                 EVQIHPDSDQISGLHGNSFHSEDE 
                 SEQ ID NO: 287 
               
               
                   
                   
                   
                 kinase, 
                   
                 IEyENQK 
               
               
                   
               
               
                 289 
                 ROS1 
                 NP_002935.2 
                 KINASE(Y); Receptor tyrosine 
                 Y363 
                 KAANMSDVSDLRIFyR 
                 SEQ ID NO: 288 
               
               
                   
                   
                   
                 kinase, 
               
               
                   
               
               
                 290 
                 TEK 
                 NP_000450.2 
                 KINASE(Y); Receptor tyrosine 
                 Y897 
                 NLLGACEHRGy 
                 SEQ ID NO: 289 
               
               
                   
                   
                   
                 kinase, 
               
               
                   
               
               
                 291 
                 TYRO3 
                 NP_006284.2 
                 KINASE(Y); Receptor tyrosine 
                 Y681 
                 KIySGDYYR 
                 SEQ ID NO: 290 
               
               
                   
                   
                   
                 kinase, 
               
               
                   
               
               
                 292 
                 DGKA 
                 NP_001336.2 
                 Kinase, lipid 
                 Y623 
                 RPHGDIyGINQALGATAK 
                 SEQ ID NO: 291 
               
               
                   
               
               
                 293 
                 PIK4CA 
                 NP_002641.1 
                 Kinase, lipid 
                 Y284 
                 yISLSEK 
                 SEQ ID NO: 292 
               
               
                   
               
               
                 294 
                 PIK3CB 
                 NP_006210.1 
                 Kinase, lipid 
                 Y962 
                 ERVPFILTyDFIHVIQQGK 
                 SEQ ID NO: 293 
               
               
                   
               
               
                 295 
                 PIK3CB 
                 NP_006210.1 
                 Kinase, lipid 
                 Y1023 
                 RHGNLFITLFALMLTAGLPELTSV 
                 SEQ ID NO: 294 
               
               
                   
                   
                   
                   
                   
                 KDIQyLK 
               
               
                   
               
               
                 296 
                 PIK3CD 
                   
                 Kinase, lipid 
                 Y484 
                 EVAPHPVyYPALEKI 
                 SEQ ID NO: 295 
               
               
                   
               
               
                 297 
                 PIK3C2B 
                 NP_002637.2 
                 Kinase, lipid 
                 Y68 
                 QNADPSLISWDEPGVDFySKPAGR 
                 SEQ ID NO: 296 
               
               
                   
               
               
                 298 
                 PIK3R1 
                 NP_852556.2 
                 Kinase, lipid 
                 Y286 
                 QAAEyREIDKR 
                 SEQ ID NOl 297 
               
               
                   
               
               
                 299 
                 EPRS 
                   
                 Ligase 
                 Y690 
                 GFFICDQPYEPVSPySCK 
                 SEQ ID NOl 298 
               
               
                   
               
               
                 300 
                 PAICS 
                 NP_006443.1 
                 Ligase 
                 Y22 
                 EVyELLDSPGK 
                 SEQ ID NO: 299 
               
               
                   
               
               
                 301 
                 OSBP 
                 NP_002547.1 
                 Lipid binding protein 
                 Y119 
                 GYQRRWFVLSNGLLSyYRSKAEMR 
                 SEQ ID NO: 300 
               
               
                   
               
               
                 302 
                 OSBP 
                 NP_002547.1 
                 Lipid binding protein 
                 Y120 
                 GYQRRWFVLSNGLLSYyRSKAEMR 
                 SEQ ID NO: 301 
               
               
                   
               
               
                 303 
                 OSBP 
                 NP_002547.1 
                 Lipid binding protein 
                 Y767 
                 EAEAMKATEDGTPYDPyKALWFER 
                 SEQ ID NO: 302 
               
               
                   
               
               
                 304 
                 DNMT1 
                 NP_001370.1 
                 Methyltransferase 
                 Y399 
                 LSIFDANESGFESyEALPQHK 
                 SEQ ID NO: 303 
               
               
                   
               
               
                 305 
                 DNMT1 
                 NP_001370.1 
                 Methyltransferase 
                 Y969 
                 KEPVDEDLyPEHYRK 
                 SEQ ID NO: 304 
               
               
                   
               
               
                 306 
                 KIAA0339 
                 NP_055527.1 
                 Methyltransferase 
                 Y748 
                 EAyHLPMPMAAEPLPSSSVSGEEA 
                 SEQ ID NO: 305 
               
               
                   
                   
                   
                   
                   
                 RLPPR 
               
               
                   
               
               
                 307 
                 DNCI2 
                 NP_001369.1 
                 Motor protein 
                 Y327 
                 TTPEyVFHCQSAVMSATFAK 
                 SEQ ID NO: 306 
               
               
                   
               
               
                 308 
                 KNS2 
                 NP_005543.2 
                 Motor protein 
                 Y449 
                 DGTSFGEyGGWYK 
                 SEQ ID NO: 307 
               
               
                   
               
               
                 309 
                 KIF20A 
                 NP_005724.1 
                 Motor protein 
                 Y869 
                 TPTCQSSTDCSPyAR 
                 SEQ ID NO: 308 
               
               
                   
               
               
                 310 
                 KLC2 
                 NP_073733.1 
                 Motor protein 
                 Y434 
                 DSAPYGEyGSWYK 
                 SEQ ID NO: 309 
               
               
                   
               
               
                 311 
                 MYO1E 
                 NP_004989.2 
                 Motor protein 
                 Y950 
                 NTTQNTGYSSGTQNANyPVR 
                 SEQ ID NO: 310 
               
               
                   
               
               
                 312 
                 MYO9B 
                 NP_004136.2 
                 Motor protein 
                 Y1683 
                 IQSHCSyTYGR 
                 SEQ ID NO: 311 
               
               
                   
               
               
                 313 
                 NYO7A 
                 NP_000251.1 
                 Motor protein 
                 Y142 
                 KIGEMPPHIFAIADNCyFNMKR 
                 SEQ ID NO: 312 
               
               
                   
               
               
                 314 
                 NY07A 
                 NP_000251.1 
                 Motor protein 
                 Y1211 
                 FVKyLRNFIHGGPPGYAPYCEER 
                 SEQ ID NO: 313 
               
               
                   
               
               
                 315 
                 MYH10 
                 NP_005955.1 
                 Motor protein 
                 Y22 
                 AVIyNPATQADWTAK 
                 SEQ ID NO: 314 
               
               
                   
               
               
                 316 
                 MYH9 
                 NP_002464.1 
                 Motor protein 
                 Y190 
                 VIQyLAYVASSHK 
                 SEQ ID NO: 315 
               
               
                   
               
               
                 317 
                 MYH9 
                 NP_002464.1 
                 Motor protein 
                 Y193 
                 KVIQYLAyVASSHK 
                 SEQ ID NO: 316 
               
               
                   
               
               
                 318 
                 SEC24C 
                   
                 Motor protein 
                 Y296 
                 ARGPQSNyGGPYPAA 
                 SEQ ID NO: 317 
               
               
                   
               
               
                 319 
                 SEC24C 
                   
                 Motor protein 
                 Y300 
                 QSNYGGPyPAAPTFG 
                 SEQ ID NO: 318 
               
               
                   
               
               
                 320 
                 TNNT2 
                 NP_000355.2 
                 Motor protein 
                 Y266 
                 yEINVLRNRINDNQKVSKTRGKAK 
                 SEQ ID NO: 319 
               
               
                   
                   
                   
                   
                   
                 VTGRWK 
               
               
                   
               
               
                 321 
                 TUBA6 
                 NP_116093.1 
                 Motor protein 
                 Y432 
                 ALEKDyEEVGADSADGEDEGEE 
                 SEQ ID NO: 320 
               
               
                   
               
               
                 322 
                 ALDH9A1 
                 NP_000687.2 
                 Oxidoreductase 
                 Y476 
                 VTIEyYSQLK 
                 SEQ ID NO: 321 
               
               
                   
               
               
                 323 
                 AKR7A2 
                 NP_003680.2 
                 Oxidoreductase 
                 Y223 
                 FyAYNPLAGGLLTGKYKYEDK 
                 SEQ ID NO: 322 
               
               
                   
               
               
                 324 
                 AKR7A2 
                 NP_003680.2 
                 Oxidoreductase 
                 Y225 
                 LGCQDAFPEVyDK 
                 SEQ ID NO: 323 
               
               
                   
               
               
                 325 
                 DHCR24 
                 NP_055577.1 
                 Oxidoreductase 
                 Y507 
                 LGCQDAFPEVyDK 
                 SEQ ID NO: 324 
               
               
                   
               
               
                 326 
                 GPD1 
                 NP_005267.2 
                 Oxidoreductase 
                 Y326 
                 LQGPETARELYSILQHKGLVDKFP 
                 SEQ ID NO: 325 
               
               
                   
                   
                   
                   
                   
                 LFMAVyK 
               
               
                   
               
               
                 327 
                 HSD17B2 
                 NP_002144.1 
                 Oxidoreductase 
                 Y232 
                 GRLVNVSSMGGGAPMERLASyGSS 
                 SEQ ID NO: 326 
               
               
                   
                   
                   
                   
                   
                 K 
               
               
                   
               
               
                 328 
                 IDH2 
                 NP_002159.2 
                 Oxidoreductase 
                 Y258 
                 AyDGRFKDIFQEIFDK 
                 SEQ ID NO: 327 
               
               
                   
               
               
                 329 
                 NDUFS7 
                 NP_077718.2 
                 Oxidoreductase 
                 Y146 
                 yVVSMGSCANGGGYYHYSYSVVR 
                 SEQ ID NO: 328 
               
               
                   
               
               
                 330 
                 NDUFS7 
                 NP_077718.2 
                 Oxidoreductase 
                 Y162 
                 YVVSMGSCANGGGYYHySYSVVR 
                 SEQ ID NO: 329 
               
               
                   
               
               
                 331 
                 ENPP3 
                 NP_005012.1 
                 Phosphatase (non-protein) 
                 Y630 
                 EyVSGFGKAMR 
                 SEQ ID NO: 330 
               
               
                   
               
               
                 332 
                 PPAP2A 
                 NP_003702.2 
                 Phosphatase (non-protein) 
                 Y168 
                 LSFySGHSSFSMYCMLFVALYLQA 
                 SEQ ID NO: 331 
               
               
                   
                   
                   
                   
                   
                 RMK 
               
               
                   
               
               
                 333 
                 PPAP2A 
                 NP_003702.2 
                 Phosphatase (non-protein) 
                 Y185 
                 LSFYSGHSSFSMYCMLFVALyLQA 
                 SEQ ID NO: 332 
               
               
                   
                   
                   
                   
                   
                 RMK 
               
               
                   
               
               
                 334 
                 INPP1 
                 NP_002185.1 
                 Phosphatase, lipid 
                 Y225 
                 WGLSyMGTNMHSLQLTISRRNGSE 
                 SEQ ID NO: 333 
               
               
                   
                   
                   
                   
                   
                 THTGNTGSEAAF 
               
               
                   
               
               
                 335 
                 INPP5D 
                 NP_001017915.1 
                 Phosphatase, lipid 
                 Y339 
                 SKDGSEDKFySHKKILQLIK 
                 SEQ ID NO: 334 
               
               
                   
               
               
                 336 
                 INPP5D 
                 NP_001017915.1 
                 Phosphatase, lipid 
                 Y1161 
                 GRDyRDNTELPHHGK 
                 SEQ ID NO: 335 
               
               
                   
               
               
                 337 
                 INPPL1 
                 NP_001558.2 
                 Phosphatase, lipid 
                 Y831 
                 SMDGyESYGECVVALK 
                 SEQ ID NO: 336 
               
               
                   
               
               
                 338 
                 INPPL1 
                   
                 Phosphatase, lipid 
                 Y1135 
                 KTLSEVDyAPAGPAR 
                 SEQ ID NO: 337 
               
               
                   
               
               
                 339 
                 INPPL1 
                   
                 Phosphatase, lipid 
                 Y1162 
                 PRGLPSDyGRPLSFP 
                 SEQ ID NO: 338 
               
               
                   
               
               
                 340 
                 PPM1G 
                 NP_002698.1 
                 PHOSPHATASE; Phosphatase 
                 Y364 
                 ALDMSyDHKPEDEVELARIK 
                 SEQ ID NO: 339 
               
               
                   
               
               
                 341 
                 PPP1R12A 
                 NP_002471.1 
                 PHOSPHATASE; Protein 
                 Y549 
                 NSSVNEGSTyHK 
                 SEQ ID NO: 340 
               
               
                   
                   
                   
                 phosphatase, regulatory 
               
               
                   
                   
                   
                 subunit; Protein phosphatase, 
               
               
                   
                   
                   
                 dual-specificity 
               
               
                   
               
               
                 342 
                 PPP1R12A 
                 NP_002471.1 
                 PHOSPHATASE; Protein 
                 Y762 
                 YSRTyDETYQR 
                 SEQ ID NO: 341 
               
               
                   
                   
                   
                 phosphatase, regulatory 
               
               
                   
                   
                   
                 subunit; Protein phosphatase, 
               
               
                   
                   
                   
                 dual-specificity 
               
               
                   
               
               
                 343 
                 PPP1CB 
                 NP_002700.1 
                 PHOSPHATASE; Protein 
                 Y306 
                 YQyGGLNSGRPVTPPR 
                 SEQ ID NO: 342 
               
               
                   
                   
                   
                 phosphatase, Ser/Thr 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 344 
                 PTPN18 
                 NP_055184.2 
                 PHOSPHATASE; Protein 
                 Y314 
                 ENCAPLyDDALFLR 
                 SEQ ID NO: 343 
               
               
                   
                   
                   
                 phosphatase, tyrosine 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 345 
                 PTPN7 
                 NP_002823.2 
                 PHOSPHATASE; Protein 
                 Y149 
                 AQSQEDGDyINANYIR 
                 SEQ ID NO: 344 
               
               
                   
                   
                   
                 phosphatase, tyrosine 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 346 
                 PTPN7 
                 NP_002823.2 
                 PHOSPHATASE; Protein 
                 Y154 
                 AQSQEDGDYINANyIR 
                 SEQ ID NO: 345 
               
               
                   
                   
                   
                 phosphatase, tyrosine 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 347 
                 PTPN6 
                   
                 PHOSPHATASE; Protein 
                 Y543 
                 SEYGNITyPPAMKNA 
                 SEQ ID NO: 346 
               
               
                   
                   
                   
                 phosphatase, tyrosine 
               
               
                   
                   
                   
                 (non-receptor) 
               
               
                   
               
               
                 348 
                 PTPRC 
                 NP_002829.2 
                 PHOSPHATASE; Receptor 
                 Y705 
                 VELSEINGDAGSNyINASYIDGFK 
                 SEQ ID NO: 347 
               
               
                   
                   
                   
                 protein phosphatase, tyrosine 
                   
                 EPR 
               
               
                   
               
               
                 349 
                 PRPRC 
                 NP_002829.2 
                 PHOSPHATASE; Receptor 
                 Y710 
                 VELSEINGDAGSNYINASyIDGFK 
                 SEQ ID NO: 348 
               
               
                   
                   
                   
                 protein phosphatase, tyrosine 
                   
                 EPR 
               
               
                   
               
               
                 350 
                 PTPRB 
                 NP_002828.2 
                 PHOSPHATASE; Receptor 
                 Y873 
                 VFPPFHLVNTATEyR 
                 SEQ ID NO: 349 
               
               
                   
                   
                   
                 protein phosphatase, tyrosine 
               
               
                   
               
               
                 351 
                 PDE2A 
                 NP_002590.1 
                 Phosphodiesterase 
                 Y920 
                 GLPSNNSLDFLDEEyEVPDLDGTR 
                 SEQ ID NO: 350 
               
               
                   
                   
                   
                   
                   
                 APINGCCSLDAE 
               
               
                   
               
               
                 352 
                 PDE8A 
                 NP_002596.1 
                 Phosphodiesterase 
                 Y194 
                 RYVENPNIMACyNELLQLEFGEVR 
                 SEQ ID NO: 351 
               
               
                   
                   
                   
                   
                   
                 SQLKLR 
               
               
                   
               
               
                 353 
                 PLCG1 
                 NP_002651.2 
                 Phospholipase 
                 Y481 
                 KLAEGSAYEEVPTSMMySENDISN 
                 SEQ ID NO: 352 
               
               
                   
                   
                   
                   
                   
                 SIK 
               
               
                   
               
               
                 354 
                 PLCG2 
                 NP_002652.1 
                 Phospholipase 
                 Y100 
                 AVRQKEDCCFTILyGTQFVLSTLS 
                 SEQ ID NO: 353 
               
               
                   
                   
                   
                   
                   
                 LAADSK 
               
               
                   
               
               
                 355 
                 PLCG2 
                 NP_002652.1 
                 Phospholipase 
                 Y680 
                 KREGSDSyAITFR 
                 SEQ ID NO: 354 
               
               
                   
               
               
                 356 
                 PLD1 
                 NP_002653.1 
                 Phospholipase 
                 Y757 
                 SAADWSAGIKyHEESIHAAYVHVI 
                 SEQ ID NO: 355 
               
               
                   
                   
                   
                   
                   
                 ENSR 
               
               
                   
               
               
                 357 
                 PLD1 
                 NP_002653.1 
                 Phospholipase 
                 Y766 
                 SAADWSAGIKYHEESIHAAyVHVI 
                 SEQ ID NO: 356 
               
               
                   
                   
                   
                   
                   
                 ENSR 
               
               
                   
               
               
                 358 
                 PLD2 
                   
                 Phospholipase 
                 Y580 
                 TPIYPyLLPK 
                 SEQ ID NO: 357 
               
               
                   
               
               
                 359 
                 PLCG2 
                   
                 Phospholipase 
                 Y1264 
                 VSNSKFyS 
                 SEQ ID NO: 358 
               
               
                   
               
               
                 360 
                 CTSK 
                   
                 Protease (non-proteasomal) 
                 Y307 
                 ENWGNKGyILMARNK 
                 SEQ ID NO: 359 
               
               
                   
               
               
                 361 
                 FAP 
                 NP_004451.2 
                 Protease (non-proteasomal) 
                 Y374 
                 DGyKHIHYIKDTVENAIQITSGK 
                 SEQ ID NO: 360 
               
               
                   
               
               
                 362 
                 FAP 
                 NP_004451.2 
                 Protease (non-proteasomal) 
                 Y379 
                 DGYKHIHyIKDTVENAIQITSGK 
                 SEQ ID NO: 361 
               
               
                   
               
               
                 363 
                 MMP9 
                 NP_004985.2 
                 Protease (non-proteasomal) 
                 Y54 
                 QLAEEYLYRYGyTRVAEMR 
                 SEQ ID NO: 362 
               
               
                   
               
               
                 364 
                 PSMA5 
                 NP_002781.2 
                 Protease (proteasomal 
                 Y8 
                 SEyDRGVNTFSPEGR 
                 SEQ ID NO: 363 
               
               
                   
                   
                   
                 subunit) 
               
               
                   
               
               
                 365 
                 PSMB4 
                 NP_002787.2 
                 Protease (proteasomal 
                 Y107 
                 VNNSTMLGASGDYADFQyLK 
                 SEQ ID NO: 364 
               
               
                   
                   
                   
                 subunit) 
               
               
                   
               
               
                 366 
                 LEPR 
                 NP_001003679.1 
                 Receptor, cytokine 
                 Y795 
                 yYIHDHFIPIEK 
                 SEQ ID NO: 365 
               
               
                   
               
               
                 367 
                 LEPR 
                 NP_001003679.1 
                 Receptor, cytokine 
                 Y796 
                 YyIHDHFIPIEK 
                 SEQ ID NO: 366 
               
               
                   
               
               
                 368 
                 MPL 
                   
                 Receptor, cytokine 
                 Y591 
                 SSQAQMDyRRPQPSC 
                 SEQ ID NO: 367 
               
               
                   
               
               
                 369 
                 ADORA2B 
                 NP_000667.1 
                 Receptor, GPCR 
                 Y113 
                 yKSLVTGTRARGVIAVLWVLAFGI 
                 SEQ ID NO: 368 
               
               
                   
                   
                   
                   
                   
                 GLTPFLGWNSK 
               
               
                   
               
               
                 370 
                 BAI3 
                 NP_001695.1 
                 Receptor, GPCR 
                 Y1419 
                 ySDLDFEKVMHTRK 
                 SEQ ID NO: 369 
               
               
                   
               
               
                 371 
                 SSTR4 
                 NP_001043.1 
                 Receptor, GPCR 
                 Y347 
                 CCLLEGAGGAEEEPLDYyATALK 
                 SEQ ID NO: 370 
               
               
                   
               
               
                 372 
                 TSHR 
                 NP_000360.2 
                 Receptor, GPCR 
                 Y643 
                 MAVLIFTDFICMAPISFyALSAIL 
                 SEQ ID NO: 371 
               
               
                   
                   
                   
                   
                   
                 NKPLITVSNSK 
               
               
                   
               
               
                 373 
                 PROCR 
                 NP_006395.2 
                 Receptor, misc. 
                 Y171 
                 PERALWQADTQVTSGVVTFTLQQL 
                 SEQ ID NO: 372 
               
               
                   
                   
                   
                   
                   
                 NAyNRTR 
               
               
                   
               
               
                 374 
                 CD19 
                 NP_001761.3 
                 Receptor, misc. 
                 Y508 
                 GILyAAPQLR 
                 SEQ ID NO: 373 
               
               
                   
               
               
                 375 
                 CD3G 
                 NP_000064.1 
                 Receptor, misc. 
                 Y160 
                 ASDKQTLLPNDQLyQPLKDREDDQ 
                 SEQ ID NO: 374 
               
               
                   
                   
                   
                   
                   
                 YSHLQGNQLR 
               
               
                   
               
               
                 376 
                 CD3G 
                 NP_000064.1 
                 Receptor, misc. 
                 Y171 
                 ASDKQTLLPNDQLYQPLKDREDDQ 
                 SEQ ID NO: 375 
               
               
                   
                   
                   
                   
                   
                 ySHLQGNQLR 
               
               
                   
               
               
                 377 
                 CD79B 
                 NP_000617.1 
                 Receptor, misc. 
                 Y196 
                 AGMEEDHTyEGLDIDQTATYEDIV 
                 SEQ ID NO: 376 
               
               
                   
                   
                   
                   
                   
                 TLR 
               
               
                   
               
               
                 378 
                 CD79B 
                 NP_000617.1 
                 Receptor, misc. 
                 Y207 
                 AGMEEDHTYEGLDIDQTATyEDIV 
                 SEQ ID NO: 377 
               
               
                   
                   
                   
                   
                   
                 TLR 
               
               
                   
               
               
                 379 
                 CR2 
                 NP_001006659.1 
                 Receptor, misc. 
                 Y108 
                 YSSCPEPIVPGGYKIRGSTPyR 
                 SEQ ID NO: 378 
               
               
                   
               
               
                 380 
                 GCER1G 
                   
                 Receptor, misc. 
                 Y65 
                 YEKSDGVyTGLSTRN 
                 SEQ ID NO: 379 
               
               
                   
               
               
                 381 
                 KIR3DL2 
                 NP_006728.1 
                 Receptor, misc. 
                 Y428 
                 TPLTDTSVyTELPNAEPR 
                 SEQ ID NO: 380 
               
               
                   
               
               
                 382 
                 P2RY2 
                 NP_002555.2 
                 Receptor, misc. 
                 Y118 
                 FLFYTNLyCSILFLTCISVHR 
                 SEQ ID NO: 381 
               
               
                   
               
               
                 383 
                 ARNT 
                 NP_001659.1 
                 Receptor, nuclear 
                 Y561 
                 FSEIyHNINADQSK 
                 SEQ ID NO: 382 
               
               
                   
               
               
                 384 
                 PPARA 
                 NP_001001928.1 
                 Receptor, nuclear 
                 Y136 
                 LKLVyDKCDRSCKIQKKNR 
                 SEQ ID NO: 383 
               
               
                   
               
               
                 385 
                 XPO7 
                 NP_055839.2 
                 Receptor, protein 
                 Y883 
                 LLLSIPHSDLLDyPK 
                 SEQ ID NO: 384 
               
               
                   
                   
                   
                 translocating 
               
               
                   
               
               
                 386 
                 RANBP5 
                 NP_002262.3 
                 Receptor, protein 
                 Y838 
                 RQDEDyDEQVEESLQDEDDNDVYI 
                 SEQ ID NO: 385 
               
               
                   
                   
                   
                 translocating 
                   
                 LTK 
               
               
                   
               
               
                 387 
                 CUTL1 
                 NP_001904.2 
                 Transcription factor 
                 Y594 
                 KyLSLSPWDKATLSMGRLVLSNKM 
                 SEQ ID NO: 386 
               
               
                   
                   
                   
                   
                   
                 AR 
               
               
                   
               
               
                 388 
                 CEBPZ 
                 NP_005751.2 
                 Transcription factor 
                 Y544 
                 yYTALYRKMLDPGLMTCSKQAMFL 
                 SEQ ID NO: 387 
               
               
                   
                   
                   
                   
                   
                 NLVYKSLK 
               
               
                   
               
               
                 389 
                 CEBPZ 
                 NP_005751.2 
                 Transcription factor 
                 Y545 
                 YyTALYRKMLDPGLMTCSKQAMFL 
                 SEQ ID NO: 388 
               
               
                   
                   
                   
                   
                   
                 NLVYKSLK 
               
               
                   
               
               
                 390 
                 CEBPZ 
                 NP_005751.2 
                 Transcription factor 
                 Y549 
                 YYTALyRKMLDPGLMTCSKQAMFL 
                 SEQ ID NO: 389 
               
               
                   
                   
                   
                   
                   
                 NLVYSKSLK 
               
               
                   
               
               
                 391 
                 CEBPZ 
                 NP_005751.2 
                 Transcription factor 
                 Y571 
                 YYTALYRKMLDPGLMTCSKQAMFL 
                 SEQ ID NO: 390 
               
               
                   
                   
                   
                   
                   
                 NLVyKSLK 
               
               
                   
               
               
                 392 
                 TCF3 
                 NP_003191.1 
                 Transcription factor 
                 Y149 
                 GTSQyYPSYSGSSR 
                 SEQ ID NO: 391 
               
               
                   
               
               
                 393 
                 TEV4 
                 NP_001977.1 
                 Transcription factor 
                 Y416 
                 YYYEKGIMQKVAGERyVYK 
                 SEQ ID NO: 392 
               
               
                   
               
               
                 394 
                 FUBP1 
                 NP_003893.2 
                 Transcription factor 
                 Y60 
                 IGGDAGTSLNSNDYGyGGQK 
                 SEQ ID NO: 393 
               
               
                   
               
               
                 395 
                 FUBP3 
                 NP_003925.1 
                 Transcription factor 
                 Y81 
                 IDSIPHLNNSTPLVDPSVYGyGVQ 
                 SEQ ID NO: 394 
               
               
                   
                   
                   
                   
                   
                 K 
               
               
                   
               
               
                 396 
                 FLI1 
                 NP_002008.2 
                 Transcription factor 
                 Y222 
                 LSVKEDPSyDSVRR 
                 SEQ ID NO: 395 
               
               
                   
               
               
                 397 
                 C21orf66 
                 NP_037461.2 
                 Transcription factor 
                 Y609 
                 yYTSYKDAYIGCLPK 
                 SEQ ID NO: 396 
               
               
                   
               
               
                 398 
                 C21orf66 
                 NP_037461.2 
                 Transcription factor 
                 Y610 
                 YyTSYKDAYIGLCLPK 
                 SEQ ID NO: 397 
               
               
                   
               
               
                 399 
                 C21orf66 
                 NP_037461.2 
                 Transcription factor 
                 Y613 
                 YYTSyKDAYIGLCLPK 
                 SEQ ID NO: 398 
               
               
                   
               
               
                 400 
                 GTF2H2 
                 NP_001506.1 
                 Transcription factor 
                 Y289 
                 PSFSMAHLDGNTEPGLTLGGyFCP 
                 SEQ ID NO: 399 
               
               
                   
                   
                   
                   
                   
                 QCRAK 
               
               
                   
               
               
                 401 
                 HKR2 
                 NP_862829.1 
                 Transcription factor 
                 Y260 
                 FDLVDAYGTEPPyTYSGKR 
                 SEQ ID NO: 400 
               
               
                   
               
               
                 402 
                 HKR2 
                 NP_862829.1 
                 Transcription factor 
                 Y262 
                 FDLVDAYGREPPYTySGKR 
                 SEQ ID NO: 401 
               
               
                   
               
               
                 403 
                 IRF5 
                 NP_002191.1 
                 Transcription factor 
                 Y136 
                 IYEVCSNGPAPTDSQPPEDySFGA 
                 SEQ ID NO: 402 
               
               
                   
                   
                   
                   
                   
                 GEEEEEEEELQR 
               
               
                   
               
               
                 404 
                 LMO4 
                 NP_006760.1 
                 Transcription factor 
                 Y37 
                 CAGCGGKIADRFLLyAMDSYWHSR 
                 SEQ ID NO: 403 
               
               
                   
                   
                   
                   
                   
                 CLK 
               
               
                   
               
               
                 405 
                 LMO4 
                 NP_006760.1 
                 Transcription factor 
                 Y42 
                 CAGCGGKIADRFLLYAMDSyWHSR 
                 SEQ ID NO: 404 
               
               
                   
                   
                   
                   
                   
                 CLK 
               
               
                   
               
               
                 406 
                 MXD1 
                 NP_002348.1 
                 Transcription factor 
                 Y18 
                 MNIQMLLEAADyLERR 
                 SEQ ID NO: 405 
               
               
                   
               
               
                 407 
                 ILF3 
                 NP_036350.2 
                 Transcription factor 
                 Y867 
                 QGGYSQSNYNSPGSGQNySGPPSS 
                 SEQ ID NO: 406 
               
               
                   
                   
                   
                   
                   
                 YQSSQGGYGR 
               
               
                   
               
               
                 408 
                 NFYA 
                 NP_002496.1 
                 Transcription factor 
                 Y266 
                 IPLPGAEMLEEEPLyVNAK 
                 SEQ ID NO: 407 
               
               
                   
               
               
                 409 
                 RAI1 
                 NP_109590.3 
                 Transcription factor 
                 Y305 
                 HHAQETLHyQNLAK 
                 SEQ ID NO: 408 
               
               
                   
               
               
                 410 
                 SPEN 
                 NP_055816.2 
                 Transcription factor 
                 Y378 
                 FGKVTSVQIHGTSEERyGLVFFR 
                 SEQ ID NO: 409 
               
               
                   
               
               
                 411 
                 STAT5A 
                 NP_003143.2 
                 Transcription factor 
                 Y22 
                 QMQVLyGQHFPEIVR 
                 SEQ ID NO: 410 
               
               
                   
               
               
                 412 
                 STAT5A 
                 NP_003143.2 
                 Transcription factor 
                 Y90 
                 LGHyATQLQK 
                 SEQ ID NO: 411 
               
               
                   
               
               
                 413 
                 STAT5A 
                 NP_003143.2 
                 Transcription factor 
                 Y114 
                 HILyNEQR 
                 SEQ ID NO: 412 
               
               
                   
               
               
                 414 
                 SMAD2 
                 NP_001003652.1 
                 Transcription factor 
                 Y102 
                 GLSTPNTIDQWDTTGLySFSEQTR 
                 SEQ ID NO: 413 
               
               
                   
                   
                   
                   
                   
                 SL 
               
               
                   
               
               
                 415 
                 ETS1 
                 NP_005229.1 
                 Transcription factor 
                 Y283 
                 VPSyDSFDSEDYPAALPNHKPK 
                 SEQ ID NO: 414 
               
               
                   
               
               
                 416 
                 NSEP1 
                 NP_003550.2 
                 Transcription factor 
                 Y145 
                 YAADRNHyRR 
                 SEQ ID NO: 415 
               
               
                   
               
               
                 417 
                 NSEP1 
                 NP_004550.2 
                 Transcription factor 
                 Y158 
                 NyQQNYQNSESGEKNEGSESAPEG 
                 SEQ ID NO: 416 
               
               
                   
                   
                   
                   
                   
                 QAQQR 
               
               
                   
               
               
                 418 
                 NSEP1 
                 NP_004550.2 
                 Transcription factor 
                 Y208 
                 RPQySNPPVQGEVMEGADNQGAGE 
                 SEQ ID NO: 417 
               
               
                   
                   
                   
                   
                   
                 QGRPVR 
               
               
                   
               
               
                 419 
                 ZNF289 
                   
                 Transcription factor 
                 Y445 
                 EVDAEyEAR 
                 SEQ ID NO: 418 
               
               
                   
               
               
                 420 
                 GTF2I 
                 NP_001509.2 
                 Transcription, 
                 Y249 
                 SEDPDYyQYNIQGSHHSSEGNE 
                 SEQ ID NO: 491 
               
               
                   
                   
                   
                 coactivator/corepressor 
               
               
                   
               
               
                 421 
                 GTF2I 
                 NP_001509.2 
                 Transcription, 
                 Y879 
                 APSyLEISSMR 
                 SEQ ID NO: 420 
               
               
                   
                   
                   
                 coactivator/corepressor 
               
               
                   
               
               
                 422 
                 SKIIP 
                 NP_036377.1 
                 Transcription, 
                 Y176 
                 AADKLAPAQyIR 
                 SEQ ID NO: 421 
               
               
                   
                   
                   
                 coactivator/corepressor 
               
               
                   
               
               
                 423 
                 SKIIP 
                 NP_036377.1 
                 Transcription, 
                 Y430 
                 DMDSGFAGGEDEIyNVYDQAWR 
                 SEQ ID NO: 422 
               
               
                   
                   
                   
                 coactivator/corepressor 
               
               
                   
               
               
                 424 
                 SKIIP 
                 NP_036377.1 
                 Transcription, 
                 Y433 
                 GMDSGFAGGEDEIYNVyDQAWR 
                 SEQ ID NO: 423 
               
               
                   
                   
                   
                 coactivator/corepressor 
               
               
                   
               
               
                 425 
                 TP53BP2 
                 NP_005417.1 
                 Transcription, 
                 Y541 
                 NIySNSQGKPGSPEPE 
                 SEQ ID NO: 424 
               
               
                   
                   
                   
                 coactivator/corepressor 
               
               
                   
               
               
                 426 
                 MPST 
                 NP_001013454.1 
                 Transferase 
                 Y272 
                 LCGKPDVPIyDGSWVEW 
                 SEQ ID NO: 425 
               
               
                   
               
               
                 427 
                 AGXT 
                 NP_000021.1 
                 Transferase 
                 Y260 
                 MyHHTIPVISLYSLR 
                 SEQ ID NO: 426 
               
               
                   
               
               
                 428 
                 ASNS 
                 NP_001664.2 
                 Transferase 
                 Y428 
                 VPFLDHRFSSyYLSLPPEMRIPK 
                 SEQ ID NO: 427 
               
               
                   
               
               
                 429 
                 ASNS 
                 NP_001664.2 
                 Transferase 
                 Y429 
                 VPFLDHRFSSYyLSLPPEMRIPK 
                 SEQ ID NO: 428 
               
               
                   
               
               
                 430 
                 BCAT1 
                 NP_005495.2 
                 Transferase 
                 Y90 
                 LHyAVELGEGLK 
                 SEQ ID NO: 429 
               
               
                   
               
               
                 431 
                 CHST1 
                 NP_003645.1 
                 Transferase 
                 Y302 
                 YMLVRyEDLARNPMKK 
                 SEQ ID NO: 430 
               
               
                   
               
               
                 432 
                 GSTP1 
                 NP_000843.1 
                 Transferase 
                 Y8 
                 PPYTVVyFPVR 
                 SEQ ID NO: 431 
               
               
                   
               
               
                 433 
                 GNPAT 
                 NP_055051.1 
                 Transferase 
                 Y60 
                 HVSDLKFAMKCYTPLVyKGITPCK 
                 SEQ ID NO: 432 
               
               
                   
               
               
                 434 
                 HAS3 
                 NP_005320.2 
                 Transferase 
                 Y520 
                 TAYCQDLFSETELAFLVSGAILyG 
                 SEQ ID NO: 433 
               
               
                   
                   
                   
                   
                   
                 CY 
               
               
                   
               
               
                 435 
                 HAS3 
                 NP_005320.2 
                 Transferase 
                 Y523 
                 TAYCQDLFSETELAFLVSGAILYG 
                 SEQ ID NO: 434 
               
               
                   
                   
                   
                   
                   
                 Cy 
               
               
                   
               
               
                 436 
                 HADHB 
                 NP_000174.1 
                 Transferase 
                 Y244 
                 LEQDEyALR 
                 SEQ ID NO: 435 
               
               
                   
               
               
                 437 
                 NMT1 
                 NP_066565.1 
                 Transferase 
                 Y476 
                 LKFGIGDGNLQyYLYNKWK 
                 SEQ ID NO: 436 
               
               
                   
               
               
                 438 
                 GLANT2 
                 NP_004472.1 
                 Transferase 
                 Y177 
                 EIILVDDySNDPEDGALLGKIEKV 
                 SEQ ID NO: 437 
               
               
                   
                   
                   
                   
                   
                 RVLRNDR 
               
               
                   
               
               
                 439 
                 POLR2H 
                 NP_006223.2 
                 Transferase 
                 Y75 
                 DGTLDDGEyNPTDDRPSRADQFE 
                 SEQ ID NO: 438 
               
               
                   
               
               
                 440 
                 Shmt1 
                   
                 Transferase 
                 Y28 
                 MLSQPLKDSDAEVySIIKK 
                 SEQ ID NO: 439 
               
               
                   
               
               
                 441 
                 SHMT1 
                 NP_004160.3 
                 Transferase 
                 Y34 
                 MLAQPLKDSDVEVYNIIK 
                 SEQ ID NO: 440 
               
               
                   
               
               
                 442 
                 B4GALT5 
                 NP_004767.1 
                 Transferase 
                 Y65 
                 DNVRTIGAQVyEQVLR 
                 SEQ ID NO: 441 
               
               
                   
               
               
                 443 
                 B4GALT5 
                 NP_004767.1 
                 Transferase 
                 Y262 
                 yMYLLPYTEFFGGVSGLTVEQFR 
                 SEQ ID NO: 442 
               
               
                   
               
               
                 444 
                 B4GALT5 
                 NP_004767.1 
                 Transferase 
                 Y264 
                 MYyLLPYTEFFGGVSGLTVEQFR 
                 SEQ ID NO: 443 
               
               
                   
               
               
                 445 
                 B4GALT5 
                 NP_004767.1 
                 Transferase 
                 Y278 
                 YMYLLPyTEFFGGVSGLTVEQFR 
                 SEQ ID NO: 444 
               
               
                   
               
               
                 446 
                 EIF4EBP2 
                 NP_004087.1 
                 Translocation initiation 
                 Y34 
                 TVAISDAAQLPHDyCTTPGGTLFS 
                 SEQ ID NO: 445 
               
               
                   
                   
                   
                 complex 
                   
                 TTPGGTR 
               
               
                   
               
               
                 447 
                 eIF2A 
                 NP_114414.2 
                 Translocation initiation 
                 Y250 
                 VIASTDVDKTGASyYGEQTLHY 
                 SEQ ID NO: 446 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 448 
                 IEF4G2 
                 NP_001409.1 
                 Translocation initiation 
                 Y439 
                 SQGLSQLyHNQSQGLLSQLQGQSK 
                 SEQ ID NO: 447 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 449 
                 EIF3S8 
                 NP_003743.1 
                 Translocation initiation 
                 Y881 
                 VFDHKQGTyGGYFR 
                 SEQ ID NO: 448 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 450 
                 EIF3S6 
                 NP_001559.1 
                 Translocation initiation 
                 Y401 
                 LGHVVMGNNAVSPyQQVIEK 
                 SEQ ID NO: 449 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 451 
                 EEF1A1 
                 NP_001393.1 
                 Translocation initiation 
                 Y86 
                 FETSKYyVTIIDAPGHR 
                 SEQ ID NO: 450 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 452 
                 EEF1D 
                 NP_001951.2 
                 Translocation initiation 
                 Y26 
                 FyEQMNGPVAGASR 
                 SEQ ID NO: 451 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 453 
                 EEF2 
                 NP_001952.1 
                 Translocation initiation 
                 Y373 
                 CELLyEGPPDDEAAMGIK 
                 SEQ ID NO: 452 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 454 
                 EIF3S6IP 
                 NP_057175.1 
                 Translocation initiation 
                 Y23 
                 AAYDPYAYPSDyDMHTGDPKQDLA 
                 SEQ ID NO: 453 
               
               
                   
                   
                   
                 complex 
                   
                 YE 
               
               
                   
               
               
                 455 
                 EIF4B 
                 NP_001408.2 
                 Translocation initiation 
                 Y266 
                 YDDRGSRDyDRGYDSR 
                 SEQ ID NO: 454 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 456 
                 EIF4B 
                 NP_001408.2 
                 Translocation initiation 
                 Y270 
                 DRYDDRGSRDYDRGyDSR 
                 SEQ ID NO: 455 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 457 
                 EIF4B 
                 NP_001408.2 
                 Translocation initiation 
                 Y291 
                 DDDyRGGGDRYEDRYDRRDDR 
                 SEQ ID NO: 456 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 458 
                 EIF4B 
                 NP_001408.2 
                 Translocation initiation 
                 Y298 
                 DDDYRGGGDRyEDRYDRRDDR 
                 SEQ ID NO: 457 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 459 
                 EIF4B 
                 NP_001408.2 
                 Translocation initiation 
                 Y593 
                 SSASKyAALSVDGEDENEGEDYAE 
                 SEQ ID NO: 458 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 460 
                 IEF4B 
                 NP_001408.2 
                 Translocation initiation 
                 Y609 
                 SSASKYAALSVDGEDENEGEDyAE 
                 SEQ ID NO: 459 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 461 
                 EIF5 
                 NP_001960.2 
                 Translocation initiation 
                 Y405 
                 VVySKAASVPKVE 
                 SEQ ID NO: 460 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 462 
                 RPS10 
                 NP_001005.1 
                 Translocation initiation 
                 Y127 
                 LTRGEADRDTyRR 
                 SEQ ID NO: 461 
               
               
                   
                   
                   
                 complex 
               
               
                   
               
               
                 463 
                 TAF15 
                 NP_003478.1 
                 Translation initiation 
                 Y50 
                 SGYGQTTDSSYGQNySGY 
                 SEQ ID NO: 462 
               
               
                   
                   
                   
                 complex; RNA binding protein 
               
               
                   
               
               
                 464 
                 TAF15 
                 NP_003478.1 
                 Translation initiation 
                 Y132 
                 DQHQGSYDEQSNyDQQHDSY 
                 SEQ ID NO: 463 
               
               
                   
                   
                   
                 complex; RNA binding protein 
               
               
                   
               
               
                 465 
                 TAF15 
                 NP_003478.1 
                 Translation initiation 
                 Y139 
                 DQHQGSYDEQSNYDQQHDSySQNQ 
                 SEQ ID NO: 464 
               
               
                   
                   
                   
                 complex; RNA binding protein 
                   
                 QSY 
               
               
                   
               
               
                 466 
                 ABCD3 
                 NP_002849.1 
                 Transporter, ABC 
                 Y267 
                 LRRPIGKMTITEQKYEGEYRyVNS 
                 SEQ ID NO: 465 
               
               
                   
                   
                   
                   
                   
                 R 
               
               
                   
               
               
                 467 
                 TAP2 
                 NP_000535.3 
                 Transporter, ABC 
                 Y693 
                 LAQLQEGQDLySR 
                 SEQ ID NO: 466 
               
               
                   
               
               
                 468 
                 ATP1A2 
                 NP_000693.1 
                 Transporter, active 
                 Y9 
                 EySPAATTAENGGGKKKQKEK 
                 SEQ ID NO: 467 
               
               
                   
               
               
                 469 
                 SLC6A6 
                 NP_003034.2 
                 Transporter, active 
                 Y598 
                 VKYLLTPREPNRWAVEREGATPyN 
                 SEQ ID NO: 468 
               
               
                   
                   
                   
                   
                   
                 SR 
               
               
                   
               
               
                 470 
                 ATP6V0A2 
                 NP_036595.2 
                 Transporter, active 
                 Y149 
                 NVEFEPTyEEFPSLESDSLLDYSC 
                 SEQ ID NO: 469 
               
               
                   
                   
                   
                   
                   
                 MQR 
               
               
                   
               
               
                 471 
                 ATP6V0A2 
                 NP_036595.2 
                 Transporter, active 
                 Y163 
                 NVEFEPTYEEFPSLESDSLLDySC 
                 SEQ ID NO: 470 
               
               
                   
                   
                   
                   
                   
                 MQR 
               
               
                   
               
               
                 472 
                 SLC1A6 
                 NP_005062.1 
                 Transporter, facilitator 
                 Y88 
                 yFSFPGELLMRMLQMLVLPLIVSS 
                 SEQ ID NO: 471 
               
               
                   
                   
                   
                   
                   
                 LVTGMASLDNK 
               
               
                   
               
               
                 473 
                 SLC1A4 
                 NP_003029.2 
                 Transporter, facilitator 
                 Y10 
                 SNETNGyLDSAQAGPAAGPGAPGT 
                 SEQ ID NO: 472 
               
               
                   
                   
                   
                   
                   
                 AAGR 
               
               
                   
               
               
                 474 
                 SLC7A2 
                 NP_001008539.1 
                 Transporter, facilitator 
                 Y621 
                 DENNEEDAyPDNVHAAAEEK 
                 SEQ ID NO: 473 
               
               
                   
               
               
                 475 
                 APC 
                 NP_000029.2 
                 Tumor suppressor 
                 Y1078 
                 NQSTTYPVyTESTDDK 
                 SEQ ID NO: 474 
               
               
                   
               
               
                 476 
                 BIRC6 
                   
                 Ubiquitin conjugating system 
                 Y4260 
                 STEEQQLyWAKGTGF 
                 SEQ ID NO: 475 
               
               
                   
               
               
                 477 
                 NYCBP2 
                 NP_055872.2 
                 Ubiquitin conjugating system 
                 Y1238 
                 FSADTDILLGGLGLFGGRGEyTAK 
                 SEQ ID NO: 476 
               
               
                   
                   
                   
                   
                   
                 IK 
               
               
                   
               
               
                 478 
                 PIAS1 
                 NP_057250.1 
                 Ubiquitin conjugating system 
                 Y144 
                 LQKLPFyDLLDELIK 
                 SEQ ID NO: 477 
               
               
                   
               
               
                 479 
                 CAND2 
                 XP_371617.2 
                 Ubiquitin conjugating system 
                 Y965 
                 PSLVRDLLDDILPLLyQETK 
                 SEQ ID NO: 478 
               
               
                   
               
               
                 480 
                 USP19 
                 NP_006668.1 
                 Ubiquitin conjugating system 
                 Y853 
                 LTyARLAQLLEGYARYSVSVFQPP 
                 SEQ ID NO: 479 
               
               
                   
                   
                   
                   
                   
                 FQPGR 
               
               
                   
               
               
                 481 
                 CUL5 
                 NP_003469.2 
                 Ubiquitin conjugating system 
                 Y373 
                 FLTARDKAyKAVVNDATIFKLELP 
                 SEQ ID NO: 480 
               
               
                   
                   
                   
                   
                   
                 LKQK 
               
               
                   
               
            
           
         
       
     
     The short name for each protein in which a phosphorylation site has presently been identified is provided in Column A, and its SwissProt accession number (human) is provided Column B. The protein type/group into which each protein falls is provided in Column C. The identified tyrosine or serine residue at which phosphorylation occurs in a given protein is identified in Column D, and the amino acid sequence of the phosphorylation site encompassing the tyrosine residue is provided in Column E (lower case y=the tyrosine (identified in Column D)) at which phosphorylation occurs. Table 1 above is identical to  FIG. 2 , except that the latter includes the disease and cell type(s) in which the particular phosphorylation site was identified (Columns F and G). 
     The identification of these 480 phosphorylation sites is described in more detail in Part A below and in Example 1. 
     Definitions 
     As used herein, the following terms have the meanings indicated: 
     “Antibody” or “antibodies” refers to all types of immunoglobulins, including IgG, IgM, IgA, IgD, and IgE, including F ab  or antigen-recognition fragments thereof, including chimeric, polyclonal, and monoclonal antibodies. The term “does not bind” with respect to an antibody&#39;s binding to one phospho-form of a sequence means does not substantially react with as compared to the antibody&#39;s binding to the other phospho-form of the sequence for which the antibody is specific. 
     “Leukemia-related signaling protein” means any protein (or poly-peptide derived therefrom) enumerated in Column A of Table 1/ FIG. 2 , which is disclosed herein as being phosphorylated in one or more leukemia cell line(s). Leukemia-related signaling proteins may be tyrosine kinases, such as Flt-3 or BCR-Abl, or serine/threonine kinases, or direct substrates of such kinases, or may be indirect substrates downstream of such kinases in signaling pathways. A Leukemia-related signaling protein may also be phosphorylated in other cell lines (non-leukemic) harboring activated kinase activity. 
     “Heavy-isotope labeled peptide” (used interchangeably with AQUA peptide) means a peptide comprising at least one heavy-isotope label, which is suitable for absolute quantification or detection of a protein as described in WO/03016861, “Absolute Quantification of Proteins and Modified Forms Thereof by Multistage Mass Spectrometry” (Gygi et al.), further discussed below. 
     “Protein” is used interchangeably with polypeptide, and includes protein fragments and domains as well as whole protein. 
     “Phosphorylatable amino acid” means any amino acid that is capable of being modified by addition of a phosphate group, and includes both forms of such amino acid. 
     “Phosphorylatable peptide sequence” means a peptide sequence comprising a phosphorylatable amino acid. 
     “Phosphorylation site-specific antibody” means an antibody that specifically binds a phosphorylatable peptide sequence/epitope only when phosphorylated, or only when not phosphorylated, respectively. The term is used interchangeably with “phospho-specific” antibody. 
     A. Identification of Novel Leukemia-Related Protein Phosphorylation Sites. 
     The nearly 480 novel Leukemia-related signaling protein phosphorylation sites disclosed herein and listed in Table 1/ FIG. 2  were discovered by employing the modified peptide isolation and characterization techniques described in “Immunoaffinity Isolation of Modified Peptides From Complex Mixtures,” U.S. Patent Publication No. 20030044848, Rush et al. (the teaching of which is hereby incorporated herein by reference, in its entirety) using cellular extracts from the following human Leukemia (AML, ALL, CML and CLL) derived cell lines and patient samples: Jurkat, K562, SEM, HT-93, CTV-1, MOLT15, CLL-9, H1993, OCL-ly3, KBM-3, UT-7, SUPT-13, MKPL-1, HU-3, M-07e, HU-3, EHEB, SU-DHL1, OCI-Iy1, DU-528, CMK, OCI-Iy8, ELF-153, OCI-Iy18, MEC-1, Karpas 299, CLL23LB4, OCI-Iy12, M01043, CLL-10, HL60, Molm 14, MV4-11, CLL-1202, EOL-1, CLL-19, CV-1, PL21; or from the following cell lines expressing activated BCR-Abl wild type and mutant kinases such as: Baf3-p210 BCR-Abl, Baf3-M351T-BCR-ABL, Baf3-E255K-BCR-Abl, Baf3-Y253F-BCR-Abl, Baf3-T315l-BCR-ABI, 3T3-v-Abl; or activated Flt3 kinase such as Baf3-FLT3 or FLT3-ITD; or JAK2 such as Baf3/Jak2; or mutant JAK2 V617F such as Baf3-V617F -JAK2, or Tyk2 such as Baf3/Tyk2; or TEL-FGFR3 such as Baf3-Tel/FGFR3; or TpoR such as Baf3/TpoR and Baf3/cc-TpoR-IV; or FGFR1 such as 293T-FGFR. The isolation and identification of phosphopeptides from these cell lines, using an immobilized general phosphotyrosine-specific antibody, or an antibody recognizing the phosphorylated motif PXpSP is described in detail in Example 1 below. In addition to the 80 previously unknown protein phosphorylation sites (tyrosine) discovered, many known phosphorylation sites were also identified (not described herein). The immunoaffinity/mass spectrometric technique described in the &#39;848 Patent Publication (the “IAP” method)—and employed as described in detail in the Examples—is briefly summarized below. 
     The IAP method employed generally comprises the following steps: (a) a proteinaceous preparation (e.g. a digested cell extract) comprising phosphopeptides from two or more different proteins is obtained from an organism; (b) the preparation is contacted with at least one immobilized general phosphotyrosine-specific antibody; (c) at least one phosphopeptide specifically bound by the immobilized antibody in step (b) is isolated; and (d) the modified peptide isolated in step (c) is characterized by mass spectrometry (MS) and/or tandem mass spectrometry (MS-MS). Subsequently, (e) a search program (e.g. Sequest) may be utilized to substantially match the spectra obtained for the isolated, modified peptide during the characterization of step (d) with the spectra for a known peptide sequence. A quantification step employing, e.g. SILAC or AQUA, may also be employed to quantify isolated peptides in order to compare peptide levels in a sample to a baseline. 
     In the IAP method as employed herein, a general phosphotyrosine-specific monoclonal antibody (commercially available from Cell Signaling Technology, Inc., Beverly, Mass., Cat #9411 (p-Tyr-100)), was used in the immunoaffinity step to isolate the widest possible number of phospho-tyrosine containing peptides from the cell extracts. 
     Extracts from the following human Leukemia cell lines (ALL, AML, CLL, CML, respectively) were employed: Jurkat, K562, SEM, HT-93, CTV-1, MOLT15, CLL-9, H1993, OCL-Iy3, KBM-3, UT-7, SUPT-13, MKPL-1, HU-3, M-07e, HU-3, EHEB, SU-DHL1, OCI-Iy1, DU-528, CMK, OCI-Iy8, ELF-153, OCI-Iy18, MEC-1, Karpas 299, CLL23LB4, OCI-Iy12, M01043, CLL-10, HL60, Molm 14, MV4-11, CLL-1202, EOL-1, CLL-19, CV-1, PL21; or from the following cell lines expressing activated BCR-Abl wild type and mutant kinases such as: Baf3-p210 BCR-Abl, Baf3-M351T-BCR-ABL, Baf3-E255K-BCR-Abl, Baf3-Y253F-BCR-Abl, Baf3-T3151-BCR-ABI, 3T3-v-Abl; or activated Flt3 kinase such as Baf3-FLT3 or FLT3-ITD; or JAK2 such as Baf3/Jak2; or mutant JAK2 V617F such as Baf3-V617F -JAK2, orTyk2 such as Baf3/Tyk2; or TEL-FGFR3 such as Baf3-Tel/FGFR3; or TpoR such as Baf3/TpoR and Baf3/cc-TpoR-IV; or FGFR1 such as 293T-FGFR. 
     As described in more detail in the Examples, lysates were prepared from these cells line and digested with trypsin after treatment with DTT and iodoacetamide to alkylate cysteine residues. Before the immunoaffinity step, peptides were pre-fractionated by reversed-phase solid phase extraction using Sep-Pak C 18  columns to separate peptides from other cellular components. The solid phase extraction cartridges were eluted with varying steps of acetonitrile. Each lyophilized peptide fraction was redissolved in PBS and treated with a phosphotyrosine antibody (P-Tyr-100, CST#9411) immobilized on protein G-Sepharose or Protein A-Sepharose. Immunoaffinity-purified peptides were eluted with 0.1% TFA and a portion of this fraction was concentrated with Stage or Zip tips and analyzed by LC-MS/MS, using a ThermoFinnigan LTQ ion trap mass spectrometer. Peptides were eluted from a 10 cm×75 μm reversed-phase column with a 45-min linear gradient of acetonitrile. MS/MS spectra were evaluated using the program Sequest with the NCBI human protein database. 
     This revealed a total of nearly 480 novel tyrosine phosphorylation sites in signaling pathways affected by kinase activation or active in leukemia cells. The identified phosphorylation sites and their parent proteins are enumerated in Table 1/ FIG. 2 . The tyrosine (human sequence) at which phosphorylation occurs is provided in Column D, and the peptide sequence encompassing the phosphorylatable tyrosine residue at the site is provided in Column E.  FIG. 2  also shows the particular type of leukemic disease (see Column G) and cell line(s) (see Column F) in which a particular phosphorylation site was discovered. 
     As a result of the discovery of these phosphorylation sites, phospho-specific antibodies and AQUA peptides for the detection of and quantification of these sites and their parent proteins may now be produced by standard methods, described below. These new reagents will prove highly useful in, e.g., studying the signaling pathways and events underlying the progression of leukemias and the identification of new biomarkers and targets for diagnosis and treatment of such diseases. 
     B. Antibodies and Cell Lines 
     Isolated phosphorylation site-specific antibodies that specifically bind a Leukemia-related signaling protein disclosed in Column A of Table 1 only when phosphorylated (or only when not phosphorylated) at the corresponding amino acid and phosphorylation site listed in Columns D and E of Table 1/ FIG. 2  may now be produced by standard antibody production methods, such as anti-peptide antibody methods, using the phosphorylation site sequence information provided in Column E of Table 1. For example, two previously unknown BCR kinase phosphorylation sites (tyrosines 58 and 231) (see Rows 225-226 of Table 1/ FIG. 2 ) are presently disclosed. Thus, antibodies that specifically bind either of these novel BCR kinase sites can now be produced, e.g. by immunizing an animal with a peptide antigen comprising all or part of the amino acid sequence encompassing the respective phosphorylated residue (e.g. a peptide antigen comprising the sequence set forth in Row 225, Column E, of Table 1 (SEQ ID NO: 224) (which encompasses the phosphorylated tyrosine at position 58 in BCR), to produce an antibody that only binds BCR kinase when phosphorylated at that site. 
     Polyclonal antibodies of the invention may be produced according to standard techniques by immunizing a suitable animal (e.g., rabbit, goat, etc.) with a peptide antigen corresponding to the Leukemia-related phosphorylation site of interest (i.e. a phosphorylation site enumerated in Column E of Table 1, which comprises the corresponding phosphorylatable amino acid listed in Column D of Table 1), collecting immune serum from the animal, and separating the polyclonal antibodies from the immune serum, in accordance with known procedures. For example, a peptide antigen corresponding to all or part of the novel ATM kinase phosphorylation site disclosed herein (SEQ ID NO: 229=EVEGTSYHESLyNALQSLRDREFYESLKYAR, encompassing phosphorylated tyrosine 2129 (see Row 230 of Table 1)) may be used to produce antibodies that only bind ATM when phosphorylated at Tyr2129. Similarly, a peptide comprising all or part of any one of the phosphorylation site sequences provided in Column E of Table 1 may employed as an antigen to produce an antibody that only binds the corresponding protein listed in Column A of Table 1 when phosphorylated (or when not phosphorylated) at the corresponding residue listed in Column D. If an antibody that only binds the protein when phosphorylated at the disclosed site is desired, the peptide antigen includes the phosphorylated form of the amino acid. Conversely, if an antibody that only binds the protein when not phosphorylated at the disclosed site is desired, the peptide antigen includes the non-phosphorylated form of the amino acid. 
     Peptide antigens suitable for producing antibodies of the invention may be designed, constructed and employed in accordance with well-known techniques. See, e.g., A NTIBODIES:  A L ABORATORY  M ANUAL,  Chapter 5, p. 75-76, Harlow &amp; Lane Eds., Cold Spring Harbor Laboratory (1988); Czernik,  Methods In Enzymology,  201: 264-283 (1991); Merrifield,  J. Am. Chem. Soc.  85: 21-49 (1962)). 
     It will be appreciated by those of skill in the art that longer or shorter phosphopeptide antigens may be employed. See Id. For example, a peptide antigen may comprise the full sequence disclosed in Column E of Table 1/ FIG. 2 , or it may comprise additional amino acids flanking such disclosed sequence, or may comprise of only a portion of the disclosed sequence immediately flanking the phosphorylatable amino acid (indicated in Column E by an uppercase “Y”). Typically, a desirable peptide antigen will comprise four or more amino acids flanking each side of the phosphorylatable amino acid and encompassing it. Polyclonal antibodies produced as described herein may be screened as further described below. 
     Monoclonal antibodies of the invention may be produced in a hybridoma cell line according to the well-known technique of Kohler and Milstein. See  Nature  265: 495-97 (1975); Kohler and Milstein,  Eur. J Immunol.  6: 511 (1976); see also, C URRENT  P ROTOCOLS IN  M OLECULAR  B IOLOGY,  Ausubel et al. Eds. (1989). Monoclonal antibodies so produced are highly specific, and improve the selectivity and specificity of diagnostic assay methods provided by the invention. For example, a solution containing the appropriate antigen may be injected into a mouse or other species and, after a sufficient time (in keeping with conventional techniques), the animal is sacrificed and spleen cells obtained. The spleen cells are then immortalized by fusing them with myeloma cells, typically in the presence of polyethylene glycol, to produce hybridoma cells. Rabbit fusion hybridomas, for example, may be produced as described in U.S. Pat. No. 5,675,063, C. Knight, Issued Oct. 7, 1997. The hybridoma cells are then grown in a suitable selection media, such as hypoxanthine-aminopterin-thymidine (HAT), and the supernatant screened for monoclonal antibodies having the desired specificity, as described below. The secreted antibody may be recovered from tissue culture supernatant by conventional methods such as precipitation, ion exchange or affinity chromatography, or the like. 
     Monoclonal Fab fragments may also be produced in  Escherichia coli  by recombinant techniques known to those skilled in the art. See, e.g., W. Huse,  Science  246:1275-81 (1989); Mullinax et al.,  Proc. Nat&#39;l Acad. Sci.  87: 8095 (1990). If monoclonal antibodies of one isotype are preferred for a particular application, particular isotypes can be prepared directly, by selecting from the initial fusion, or prepared secondarily, from a parental hybridoma secreting a monoclonal antibody of different isotype by using the sib selection technique to isolate class-switch variants (Steplewski, et al.,  Proc. Nat&#39;l. Acad. Sci.,  82: 8653 (1985); Spira et al.,  J. Immunol. Methods,  74: 307 (1984)). 
     The preferred epitope of a phosphorylation-site specific antibody of the invention is a peptide fragment consisting essentially of about 8 to 17 amino acids including the phosphorylatable tyrosine, wherein about 3 to 8 amino acids are positioned on each side of the phosphorylatable tyrosine (for example, the MELK tyrosine 438 phosphorylation site sequence disclosed in Row 244, Column E of Table 1), and antibodies of the invention thus specifically bind a target Leukemia-related signaling polypeptide comprising such epitopic sequence. Particularly preferred epitopes bound by the antibodies of the invention comprise all or part of a phosphorylatable site sequence listed in Column E of Table 1, including the phosphorylatable amino acid. 
     Included in the scope of the invention are equivalent non-antibody molecules, such as protein binding domains or nucleic acid aptamers, which bind, in a phospho-specific manner, to essentially the same phosphorylatable epitope to which the phospho-specific antibodies of the invention bind. See, e.g., Neuberger et al.,  Nature  312: 604 (1984). Such equivalent non-antibody reagents may be suitably employed in the methods of the invention further described below. 
     Antibodies provided by the invention may be any type of immunoglobulins, including IgG, IgM, IgA, IgD, and IgE, including F ab  or antigen-recognition fragments thereof. The antibodies may be monoclonal or polyclonal and may be of any species of origin, including (for example) mouse, rat, rabbit, horse, or human, or may be chimeric antibodies. See, e.g., M. Walker et al.,  Molec. ImmunoL  26: 403-11 (1989); Morrision et al.,  Proc. Nat&#39;l. Acad. Sci.  81: 6851 (1984); Neuberger et al.,  Nature  312: 604 (1984)). The antibodies may be recombinant monoclonal antibodies produced according to the methods disclosed in U.S. Pat. No. 4,474,893 (Reading) or U.S. Pat. No. 4,816,567 (Cabilly et al.) The antibodies may also be chemically constructed by specific antibodies made according to the method disclosed in U.S. Pat. No. 4,676,980 (Segel et al.) 
     The invention also provides immortalized cell lines that produce an antibody of the invention. For example, hybridoma clones, constructed as described above, that produce monoclonal antibodies to the Leukemia-related signaling protein phosphorylation sties disclosed herein are also provided. Similarly, the invention includes recombinant cells producing an antibody of the invention, which cells may be constructed by well known techniques; for example the antigen combining site of the monoclonal antibody can be cloned by PCR and single-chain antibodies produced as phage-displayed recombinant antibodies or soluble antibodies in  E. coli  (see, e.g., A NTIBODY  E NGINEERING  P ROTOCOLS,  1995, Humana Press, Sudhir Paul editor.) 
     Phosphorylation site-specific antibodies of the invention, whether polyclonal or monoclonal, may be screened for epitope and phospho-specificity according to standard techniques. See, e.g. Czemik et al.,  Methods in Enzymology,  201: 264-283 (1991). For example, the antibodies may be screened against the phospho and non-phospho peptide library by ELISA to ensure specificity for both the desired antigen (i.e. that epitope including a phosphorylation site sequence enumerated in Column E of Table 1) and for reactivity only with the phosphorylated (or non-phosphorylated) form of the antigen. Peptide competition assays may be carried out to confirm lack of reactivity with other phospho-epitopes on the given Leukemia-related signaling protein. The antibodies may also be tested by Western blotting against cell preparations containing the signaling protein, e.g. cell lines over-expressing the target protein, to confirm reactivity with the desired phosphorylated epitope/target. 
     Specificity against the desired phosphorylated epitope may also be examined by constructing mutants lacking phosphorylatable residues at positions outside the desired epitope that are known to be phosphorylated, or by mutating the desired phospho-epitope and confirming lack of reactivity. Phosphorylation-site specific antibodies of the invention may exhibit some limited cross-reactivity to related epitopes in non-target proteins. This is not unexpected as most antibodies exhibit some degree of cross-reactivity, and anti-peptide antibodies will often cross-react with epitopes having high homology to the immunizing peptide. See, e.g., Czernik, supra. Cross-reactivity with non-target proteins is readily characterized by Western blotting alongside markers of known molecular weight. Amino acid sequences of cross-reacting proteins may be examined to identify sites highly homologous to the Leukemia-related signaling protein epitope for which the antibody of the invention is specific. 
     In certain cases, polyclonal antisera may exhibit some undesirable general cross-reactivity to phosphotyrosine itself, which may be removed by further purification of antisera, e.g. over a phosphotyramine column. Antibodies of the invention specifically bind their target protein (i.e. a protein listed in Column A of Table 1) only when phosphorylated (or only when not phosphorylated, as the case may be) at the site disclosed in corresponding Columns D/E, and do not (substantially) bind to the other form (as compared to the form for which the antibody is specific). 
     Antibodies may be further characterized via immunohistochemical (IHC) staining using normal and diseased tissues to examine Leukemia-related phosphorylation and activation status in diseased tissue. IHC may be carried out according to well-known techniques. See, e.g., A NTIBODIES:  A L ABORATORY  M ANUAL,  Chapter 10, Harlow &amp; Lane Eds., Cold Spring Harbor Laboratory (1988). Briefly, paraffin-embedded tissue (e.g. tumor tissue) is prepared for immunohistochemical staining by deparaffinizing tissue sections with xylene followed by ethanol; hydrating in water then PBS; unmasking antigen by heating slide in sodium citrate buffer; incubating sections in hydrogen peroxide; blocking in blocking solution; incubating slide in primary antibody and secondary antibody; and finally detecting using ABC avidin/biotin method according to manufacturer&#39;s instructions. 
     Antibodies may be further characterized by flow cytometry carried out according to standard methods. See Chow et al.,  Cytometry  ( Communications in Clinical Cytometry ) 46: 72-78 (2001). Briefly and by way of example, the following protocol for cytometric analysis may be employed: samples may be centrifuged on Ficoll gradients to remove erythrocytes, and cells may then be fixed with 2% paraformaldehyde for 10 minutes at 37° C. followed by permeabilization in 90% methanol for 30 minutes on ice. Cells may then be stained with the primary phosphorylation-site specific antibody of the invention (which detects a Leukemia-related signal transduction protein enumerated in Table 1), washed and labeled with a fluorescent-labeled secondary antibody. Additional fluorochrome-conjugated marker antibodies (e.g. CD45, CD34) may also be added at this time to aid in the subsequent identification of specific hematopoietic cell types. The cells would then be analyzed on a flow cytometer (e.g. a Beckman Coulter FC500) according to the specific protocols of the instrument used. 
     Antibodies of the invention may also be advantageously conjugated to fluorescent dyes (e.g. Alexa488, PE) for use in multi-parametric analyses along with other signal transduction (phospho-CrkL, phospho-Erk 1/2) and/or cell marker (CD34) antibodies. 
     Phosphorylation-site specific antibodies of the invention specifically bind to a human Leukemia-related signal transduction protein or polypeptide only when phosphorylated at a disclosed site, but are not limited only to binding the human species, per se. The invention includes antibodies that also bind conserved and highly homologous or identical phosphorylation sites in respective Leukemia-related proteins from other species (e.g. mouse, rat, monkey, yeast), in addition to binding the human phosphorylation site. Highly homologous or identical sites conserved in other species can readily be identified by standard sequence comparisons, such as using BLAST, with the human Leukemia-related signal transduction protein phosphorylation sites disclosed herein. 
     C. Heavy-Isotope Labeled Peptides (AQUA Peptides). 
     The novel Leukemia-related signaling protein phosphorylation sites disclosed herein now enable the production of corresponding heavy-isotope labeled peptides for the absolute quantification of such signaling proteins (both phosphorylated and not phosphorylated at a disclosed site) in biological samples. The production and use of AQUA peptides for the absolute quantification of proteins (AQUA) in complex mixtures has been described. See WO/03016861, “Absolute Quantification of Proteins and Modified Forms Thereof by Multistage Mass Spectrometry,” Gygi et al. and also Gerber et al.  Proc. Nati. Acad. Sci. U.S.A.  100: 6940-5 (2003) (the teachings of which are hereby incorporated herein by reference, in their entirety). 
     The AQUA methodology employs the introduction of a known quantity of at least one heavy-isotope labeled peptide standard (which has a unique signature detectable by LC-SRM chromatography) into a digested biological sample in order to determine, by comparison to the peptide standard, the absolute quantity of a peptide with the same sequence and protein modification in the biological sample. Briefly, the AQUA methodology has two stages: peptide internal standard selection and validation and method development; and implementation using validated peptide internal standards to detect and quantify a target protein in sample. The method is a powerful technique for detecting and quantifying a given peptide/protein within a complex biological mixture, such as a cell lysate, and may be employed, e.g., to quantify change in protein phosphorylation as a result of drug treatment, or to quantify differences in the level of a protein in different biological states. 
     Generally, to develop a suitable internal standard, a particular peptide (or modified peptide) within a target protein sequence is chosen based on its amino acid sequence and the particular protease to be used to digest. The peptide is then generated by solid-phase peptide synthesis such that one residue is replaced with that same residue containing stable isotopes ( 13 C,  15 N). The result is a peptide that is chemically identical to its native counterpart formed by proteolysis, but is easily distinguishable by MS via a 7-Da mass shift. A newly synthesized AQUA internal standard peptide is then evaluated by LC-MS/MS. This process provides qualitative information about peptide retention by reverse-phase chromatography, ionization efficiency, and fragmentation via collision-induced dissociation. Informative and abundant fragment ions for sets of native and internal standard peptides are chosen and then specifically monitored in rapid succession as a function of chromatographic retention to form a selected reaction monitoring (LC-SRM) method based on the unique profile of the peptide standard. 
     The second stage of the AQUA strategy is its implementation to measure the amount of a protein or modified protein from complex mixtures. Whole cell lysates are typically fractionated by SDS-PAGE gel electrophoresis, and regions of the gel consistent with protein migration are excised. This process is followed by in-gel proteolysis in the presence of the AQUA peptides and LC-SRM analysis. (See Gerber et al. supra.) AQUA peptides are spiked in to the complex peptide mixture obtained by digestion of the whole cell lysate with a proteolytic enzyme and subjected to immunoaffinity purification as described above. The retention time and fragmentation pattern of the native peptide formed by digestion (e.g. trypsinization) is identical to that of the AQUA internal standard peptide determined previously; thus, LC-MS/MS analysis using an SRM experiment results in the highly specific and sensitive measurement of both internal standard and analyte directly from extremely complex peptide mixtures. Because an absolute amount of the AQUA peptide is added (e.g. 250 fmol), the ratio of the areas under the curve can be used to determine the precise expression levels of a protein or phosphorylated form of a protein in the original cell lysate. In addition, the internal standard is present during in-gel digestion as native peptides are formed, such that peptide extraction efficiency from gel pieces, absolute losses during sample handling (including vacuum centrifugation), and variability during introduction into the LC-MS system do not affect the determined ratio of native and AQUA peptide abundances. 
     An AQUA peptide standard is developed for a known phosphorylation site sequence previously identified by the IAP-LC-MS/MS method within a target protein. One AQUA peptide incorporating the phosphorylated form of the particular residue within the site may be developed, and a second AQUA peptide incorporating the non-phosphorylated form of the residue developed. In this way, the two standards may be used to detect and quantify both the phosphorylated and non-phosphorylated forms of the site in a biological sample. 
     Peptide internal standards may also be generated by examining the primary amino acid sequence of a protein and determining the boundaries of peptides produced by protease cleavage. Alternatively, a protein may actually be digested with a protease and a particular peptide fragment produced can then sequenced. Suitable proteases include, but are not limited to, serine proteases (e.g. trypsin, hepsin), metallo proteases (e.g. PUMP1), chymotrypsin, cathepsin, pepsin, thermolysin, carboxypeptidases, etc. 
     A peptide sequence within a target protein is selected according to one or more criteria to optimize the use of the peptide as an internal standard. Preferably, the size of the peptide is selected to minimize the chances that the peptide sequence will be repeated elsewhere in other non-target proteins. Thus, a peptide is preferably at least about 6 amino acids. The size of the peptide is also optimized to maximize ionization frequency. Thus, peptides longer than about 20 amino acids are not preferred. The preferred ranged is about 7 to 15 amino acids. A peptide sequence is also selected that is not likely to be chemically reactive during mass spectrometry, thus sequences comprising cysteine, tryptophan, or methionine are avoided. 
     A peptide sequence that does not include a modified region of the target region may be selected so that the peptide internal standard can be used to determine the quantity of all forms of the protein. Alternatively, a peptide internal standard encompassing a modified amino acid may be desirable to detect and quantify only the modified form of the target protein. Peptide standards for both modified and unmodified regions can be used together, to determine the extent of a modification in a particular sample (i.e. to determine what fraction of the total amount of protein is represented by the modified form). For example, peptide standards for both the phosphorylated and unphosphorylated form of a protein known to be phosphorylated at a particular site can be used to quantify the amount of phosphorylated form in a sample. 
     The peptide is labeled using one or more labeled amino acids (i.e. the label is an actual part of the peptide) or less preferably, labels may be attached after synthesis according to standard methods. Preferably, the label is a mass-altering label selected based on the following considerations: The mass should be unique to shift fragment masses produced by MS analysis to regions of the spectrum with low background; the ion mass signature component is the portion of the labeling moiety that preferably exhibits a unique ion mass signature in MS analysis; the sum of the masses of the constituent atoms of the label is preferably uniquely different than the fragments of all the possible amino acids. As a result, the labeled amino acids and peptides are readily distinguished from unlabeled ones by the ion/mass pattern in the resulting mass spectrum. Preferably, the ion mass signature component imparts a mass to a protein fragment that does not match the residue mass for any of the 20 natural amino acids. 
     The label should be robust under the fragmentation conditions of MS and not undergo unfavorable fragmentation. Labeling chemistry should be efficient under a range of conditions, particularly denaturing conditions, and the labeled tag preferably remains soluble in the MS buffer system of choice. The label preferably does not suppress the ionization efficiency of the protein and is not chemically reactive. The label may contain a mixture of two or more isotopically distinct species to generate a unique mass spectrometric pattern at each labeled fragment position. Stable isotopes, such as  2 H,  13 C,  15 N,  17 O,  18 O, or  34 S, are among preferred labels. Pairs of peptide internal standards that incorporate a different isotope label may also be prepared. Preferred amino acid residues into which a heavy isotope label may be incorporated include leucine, proline, valine, and phenylalanine. 
     Peptide internal standards are characterized according to their mass-to-charge (m/z) ratio, and preferably, also according to their retention time on a chromatographic column (e.g. an HPLC column). Internal standards that co-elute with unlabeled peptides of identical sequence are selected as optimal internal standards. The internal standard is then analyzed by fragmenting the peptide by any suitable means, for example by collision-induced dissociation (CID) using, e.g., argon or helium as a collision gas. The fragments are then analyzed, for example by multi-stage mass spectrometry (MS n ) to obtain a fragment ion spectrum, to obtain a peptide fragmentation signature. Preferably, peptide fragments have significant differences in m/z ratios to enable peaks corresponding to each fragment to be well separated, and a signature that is unique for the target peptide is obtained. If a suitable fragment signature is not obtained at the first stage, additional stages of MS are performed until a unique signature is obtained. 
     Fragment ions in the MS/MS and MS 3  spectra are typically highly specific for the peptide of interest, and, in conjunction with LC methods, allow a highly selective means of detecting and quantifying a target peptide/protein in a complex protein mixture, such as a cell lysate, containing many thousands or tens of thousands of proteins. Any biological sample potentially containing a target protein/peptide of interest may be assayed. Crude or partially purified cell extracts are preferably employed. Generally, the sample has at least 0.01 mg of protein, typically a concentration of 0.1-10 mg/mL, and may be adjusted to a desired buffer concentration and pH. 
     A known amount of a labeled peptide internal standard, preferably about 10 femtomoles, corresponding to a target protein to be detected/quantified is then added to a biological sample, such as a cell lysate. The spiked sample is then digested with one or more protease(s) for a suitable time period to allow digestion. A separation is then performed (e.g. by HPLC, reverse-phase HPLC, capillary electrophoresis, ion exchange chromatography, etc.) to isolate the labeled internal standard and its corresponding target peptide from other peptides in the sample. Microcapillary LC is a preferred method. 
     Each isolated peptide is then examined by monitoring of a selected reaction in the MS. This involves using the prior knowledge gained by the characterization of the peptide internal standard and then requiring the MS to continuously monitor a specific ion in the MS/MS or MS n  spectrum for both the peptide of interest and the internal standard. After elution, the area under the curve (AUC) for both peptide standard and target peptide peaks are calculated. The ratio of the two areas provides the absolute quantification that can be normalized for the number of cells used in the analysis and the protein&#39;s molecular weight, to provide the precise number of copies of the protein per cell. Further details of the AQUA methodology are described in Gygi et al., and Gerber et al. supra. 
     In accordance with the present invention, AQUA internal peptide standards (heavy-isotope labeled peptides) may now be produced, as described above, for any of the nearly 480 novel Leukemia-related signaling protein phosphorylation sites disclosed herein (see Table 1/ FIG. 2 ). Peptide standards for a given phosphorylation site (e.g. the tyrosine 187 in BLK—see Row 271 of Table 1) may be produced for both the phosphorylated and non-phosphorylated forms of the site (e.g. see BLK site sequence in Column E, Row 271 of Table 1 (SEQ ID NO: 270) and such standards employed in the AQUA methodology to detect and quantify both forms of such phosphorylation site in a biological sample. 
     AQUA peptides of the invention may comprise all, or part of, a phosphorylation site peptide sequence disclosed herein (see Column E of Table 1/ FIG. 2 ). In a preferred embodiment, an AQUA peptide of the invention comprises a phosphorylation site sequence disclosed herein in Table 1/ FIG. 2 . For example, an AQUA peptide of the invention for detection/quantification of SYK kinase when phosphorylated at tyrosine Y296 may comprise the sequence IKSySFPKPGHR (y=phosphotyrosine), which comprises phosphorylatable tyrosine 296 (see Row 274, Column E; (SEQ ID NO: 273)). Heavy-isotope labeled equivalents of the peptides enumerated in Table 1/ FIG. 2  (both in phosphorylated and unphosphorylated form) can be readily synthesized and their unique MS and LC-SRM signature determined, so that the peptides are validated as AQUA peptides and ready for use in quantification experiments. 
     The phosphorylation site peptide sequences disclosed herein (see Column E of Table 1/ FIG. 2 ) are particularly well suited for development of corresponding AQUA peptides, since the IAP method by which they were identified (see Part A above and Example 1) inherently confirmed that such peptides are in fact produced by enzymatic digestion (trypsinization) and are in fact suitably fractionated/ionized in MS/MS. Thus, heavy-isotope labeled equivalents of these peptides (both in phosphorylated and unphosphorylated form) can be readily synthesized and their unique MS and LC-SRM signature determined, so that the peptides are validated as AQUA peptides and ready for use in quantification experiments. 
     Accordingly, the invention provides heavy-isotope labeled peptides (AQUA peptides) for the detection and/or quantification of any of the Leukemia-related phosphorylation sites disclosed in Table 1/ FIG. 2  (see Column E) and/or their corresponding parent proteins/polypeptides (see Column A). A phosphopeptide sequence comprising any of the phosphorylation sequences listed in Table 1 may be considered a preferred AQUA peptide of the invention. For example, an AQUA peptide comprising the sequence VMNATAyGISK (SEQ ID NO:285) (where y may be either phosphotyrosine or tyrosine, and where V=labeled valine (e.g.  14 C)) is provided for the quantification of phosphorylated (or non-phosphorylated) FLT3 kinase (Tyr630) in a biological sample (see Row 286 of Table 1, tyrosine 630 being the phosphorylatable residue within the site). However, it will be appreciated that a larger AQUA peptide comprising a disclosed phosphorylation site sequence (and additional residues downstream or upstream of it) may also be constructed. Similarly, a smaller AQUA peptide comprising less than all of the residues of a disclosed phosphorylation site sequence (but still comprising the phosphorylatable residue enumerated in Column D of Table 1/ FIG. 2 ) may alternatively be constructed. Such larger or shorter AQUA peptides are within the scope of the present invention, and the selection and production of preferred AQUA peptides may be carried out as described above (see Gygi et al., Gerber et al. supra.). 
     Certain particularly preferred subsets of AQUA peptides provided by the invention are described above (corresponding to particular protein types/groups in Table 1, for example, Tyrosine Protein Kinases or Protein Phosphatases). Example 4 is provided to further illustrate the construction and use, by standard methods described above, of exemplary AQUA peptides provided by the invention. For example, the above-described AQUA peptides corresponding to the both the phosphorylated and non-phosphorylated forms of the disclosed FLT3 kinase tyrosine 630 phosphorylation site (see Row 286 of Table 1/ FIG. 2 ) may be used to quantify the amount of phosphorylated FLT3 (Tyr630) in a biological sample, e.g. a tumor cell sample (or a sample before or after treatment with a test drug). 
     AQUA peptides of the invention may also be employed within a kit that comprises one or multiple AQUA peptide(s) provided herein (for the quantification of a Leukemia-related signal transduction protein disclosed in Table 1/ FIG. 2 ), and, optionally, a second detecting reagent conjugated to a detectable group. For example, a kit may include AQUA peptides for both the phosphorylated and non-phosphorylated form of a phosphorylation site disclosed herein. The reagents may also include ancillary agents such as buffering agents and protein stabilizing agents, e.g., polysaccharides and the like. The kit may further include, where necessary, other members of the signal-producing system of which system the detectable group is a member (e.g., enzyme substrates), agents for reducing background interference in a test, control reagents, apparatus for conducting a test, and the like. The test kit may be packaged in any suitable manner, typically with all elements in a single container along with a sheet of printed instructions for carrying out the test. 
     AQUA peptides provided by the invention will be highly useful in the further study of signal transduction anomalies underlying cancer, including leukemias, and in identifying diagnostic/bio-markers of these diseases, new potential drug targets, and/or in monitoring the effects of test compounds on Leukemia-related signal transduction proteins and pathways. 
     D. Immunoassay Formats 
     Antibodies provided by the invention may be advantageously employed in a variety of standard immunological assays (the use of AQUA peptides provided by the invention is described separately above). Assays may be homogeneous assays or heterogeneous assays. In a homogeneous assay the immunological reaction usually involves a phosphorylation-site specific antibody of the invention), a labeled analyte, and the sample of interest. The signal arising from the label is modified, directly or indirectly, upon the binding of the antibody to the labeled analyte. Both the immunological reaction and detection of the extent thereof are carried out in a homogeneous solution. Immunochemical labels that may be employed include free radicals, radioisotopes, fluorescent dyes, enzymes, bacteriophages, coenzymes, and so forth. 
     In a heterogeneous assay approach, the reagents are usually the specimen, a phosphorylation-site specific antibody of the invention, and suitable means for producing a detectable signal. Similar specimens as described above may be used. The antibody is generally immobilized on a support, such as a bead, plate or slide, and contacted with the specimen suspected of containing the antigen in a liquid phase. The support is then separated from the liquid phase and either the support phase or the liquid phase is examined for a detectable signal employing means for producing such signal. The signal is related to the presence of the analyte in the specimen. Means for producing a detectable signal include the use of radioactive labels, fluorescent labels, enzyme labels, and so forth. For example, if the antigen to be detected contains a second binding site, an antibody which binds to that site can be conjugated to a detectable group and added to the liquid phase reaction solution before the separation step. The presence of the detectable group on the solid support indicates the presence of the antigen in the test sample. Examples of suitable immunoassays are the radioimmunoassay, immunofluorescence methods, enzyme-linked immunoassays, and the like. 
     Immunoassay formats and variations thereof that may be useful for carrying out the methods disclosed herein are well known in the art. See generally E. Maggio, Enzyme-Immunoassay, (1980) (CRC Press, Inc., Boca Raton, Fla.); see also, e.g., U.S. Pat. No. 4,727,022 (Skold et al., “Methods for Modulating Ligand-Receptor Interactions and their Application”); U.S. Pat. No. 4,659,678 (Forrest et al., “Immunoassay of Antigens”); U.S. Pat. No. 4,376,110 (David et al., “Immunometric Assays Using Monoclonal Antibodies”). Conditions suitable for the formation of reagent-antibody complexes are well described. See id. Monoclonal antibodies of the invention may be used in a “two-site” or “sandwich” assay, with a single cell line serving as a source for both the labeled monoclonal antibody and the bound monoclonal antibody. Such assays are described in U.S. Pat. No. 4,376,110. The concentration of detectable reagent should be sufficient such that the binding of a target Leukemia-related signal transduction protein is detectable compared to background. 
     Phosphorylation site-specific antibodies disclosed herein may be conjugated to a solid support suitable for a diagnostic assay (e.g., beads, plates, slides or wells formed from materials such as latex or polystyrene) in accordance with known techniques, such as precipitation. Antibodies, or other target protein or target site-binding reagents, may likewise be conjugated to detectable groups such as radiolabels (e.g.,  35 S,  125 I,  131 I), enzyme labels (e.g., horseradish peroxidase, alkaline phosphatase), and fluorescent labels (e.g., fluorescein) in accordance with known techniques. 
     Antibodies of the invention may also be optimized for use in a flow cytometry (FC) assay to determine the activation/phosphorylation status of a target Leukemia-related signal transduction protein in patients before, during, and after treatment with a drug targeted at inhibiting phosphorylation at such a protein at the phosphorylation site disclosed herein. For example, bone marrow cells or peripheral blood cells from patients may be analyzed by flow cytometry for target Leukemia-related signal transduction protein phosphorylation, as well as for markers identifying various hematopoietic cell types. In this manner, activation status of the malignant cells may be specifically characterized. Flow cytometry may be carried out according to standard methods. See, e.g. Chow et al.,  Cytometry  ( Communications in Clinical Cytometry ) 46: 72-78 (2001). Briefly and by way of example, the following protocol for cytometric analysis may be employed: fixation of the cells with 1% para-formaldehyde for 10 minutes at 37° C. followed by permeabilization in 90% methanol for 30 minutes on ice. Cells may then be stained with the primary antibody (a phospho-specific antibody of the invention), washed and labeled with a fluorescent-labeled secondary antibody. Alternatively, the cells may be stained with a fluorescent-labeled primary antibody. The cells would then be analyzed on a flow cytometer (e.g. a Beckman Coulter EPICS-XL) according to the specific protocols of the instrument used. Such an analysis would identify the presence of activated Leukemia-related signal transduction protein(s) in the malignant cells and reveal the drug response on the targeted protein. 
     Alternatively, antibodies of the invention may be employed in immunohistochemical (IHC) staining to detect differences in signal transduction or protein activity using normal and diseased tissues. IHC may be carried out according to well-known techniques. See, e.g., A NTIBODIES:  A L ABORATORY  M ANUAL,  supra. Briefly, paraffin-embedded tissue (e.g. tumor tissue) is prepared for immunohistochemical staining by deparaffinizing tissue sections with xylene followed by ethanol; hydrating in water then PBS; unmasking antigen by heating slide in sodium citrate buffer; incubating sections in hydrogen peroxide; blocking in blocking solution; incubating slide in primary antibody and secondary antibody; and finally detecting using ABC avidin/biotin method according to manufacturer&#39;s instructions. 
     Antibodies of the invention may be also be optimized for use in other clinically-suitable applications, for example bead-based multiplex-type assays, such as IGEN, Luminex™ and/or Bioplex™ assay formats, or otherwise optimized for antibody arrays formats, such as reversed-phase array applications (see, e.g. Paweletz et al.,  Oncogene  20(16): 1981-89 (2001)). Accordingly, in another embodiment, the invention provides a method for the multiplex detection of Leukemia-related protein phosphorylation in a biological sample, the method comprising utilizing two or more antibodies or AQUA peptides of the invention to detect the presence of two or more phosphorylated Leukemia-related signaling proteins enumerated in Column A of Table 1/ FIG. 2 . In one preferred embodiment, two to five antibodies or AQUA peptides of the invention are employed in the method. In another preferred embodiment, six to ten antibodies or AQUA peptides of the invention are employed, while in another preferred embodiment eleven to twenty such reagents are employed. 
     Antibodies and/or AQUA peptides of the invention may also be employed within a kit that comprises at least one phosphorylation site-specific antibody or AQUA peptide of the invention (which binds to or detects a Leukemia-related signal transduction protein disclosed in Table 1/ FIG. 2 ), and, optionally, a second antibody conjugated to a detectable group. In some embodies, the kit is suitable for multiplex assays and comprises two or more antibodies or AQUA peptides of the invention, and in some embodiments, comprises two to five, six to ten, or eleven to twenty reagents of the invention. The kit may also include ancillary agents such as buffering agents and protein stabilizing agents, e.g., polysaccharides and the like. The kit may further include, where necessary, other members of the signal-producing system of which system the detectable group is a member (e.g., enzyme substrates), agents for reducing background interference in a test, control reagents, apparatus for conducting a test, and the like. The test kit may be packaged in any suitable manner, typically with all elements in a single container along with a sheet of printed instructions for carrying out the test. 
     The following Examples are provided only to further illustrate the invention, and are not intended to limit its scope, except as provided in the claims appended hereto. The present invention encompasses modifications and variations of the methods taught herein which would be obvious to one of ordinary skill in the art. 
     EXAMPLE 1  
     Isolation of Phosphotyrosine-Containing Peptides from Extracts of Leukemia Cell Lines and Identification of Novel Phosphorylation Sites 
     In order to discover previously unknown Leukemia-related signal transduction protein phosphorylation sites, IAP isolation techniques were employed to identify phosphotyrosine- and/or phosphoserine-containing peptides in cell extracts from the following human Leukemia cell lines and patient cell lines: Jurkat, K562, SEM, HT-93, CTV-1, MOLT15, CLL-9, H1993, OCL-Iy3, KBM-3, UT-7, SUPT-13, MKPL-1, HU-3, M-07e, HU-3, EHEB, SU-DHL1, OCI-Iy1, DU-528, CMK, OCI-Iy8, ELF-153, OCI-Iy18, MEC-1, Karpas 299, CLL23LB4, OCI-Iy12, M01043, CLL-10, HL60, Molm 14, MV4-11, CLL-1202, EOL-1, CLL-19, CV-1, PL21; or from the following cell lines expressing activated BCR-Abl wild type and mutant kinases such as: Baf3-p210 BCR-Abl, Baf3-M351T-BCR-ABL, Baf3-E255K-BCR-Abl, Baf3-Y253F-BCR-Abl, Baf3-T3151-BCR-ABI, 3T3-v-Abl; or activated Flt3 kinase such as Baf3-FLT3 or FLT3-ITD; or JAK2 such as Baf3/Jak2; or mutant JAK2 V617F such as Baf3-V617F -JAK2, or Tyk2 such as Baf3/Tyk2; or TEL-FGFR3 such as Baf3-Tel/FGFR3; or TpoR such as Baf3/TpoR and Baf3/cc-TpoR-IV; or FGFR1 such as 293T-FGFR. 
     Tryptic phosphotyrosine—containing peptides were purified and analyzed from extracts of each of the 29 cell lines mentioned above, as follows. Cells were cultured in DMEM medium or RPMI 1640 medium supplemented with 10% fetal bovine serum and penicillin/streptomycin. 
     Suspension cells were harvested by low speed centrifugation. After complete aspiration of medium, cells were resuspended in 1 mL lysis buffer per 1.25×10 8  cells (20 mM HEPES pH 8.0, 9 M urea, 1 mM sodium vanadate, supplemented or not with 2.5 mM sodium pyro-phosphate, 1 mM β-glycerol-phosphate) and sonicated. 
     Sonicated cell lysates were cleared by centrifugation at 20,000×g, and proteins were reduced with DTT at a final concentration of 4.1 mM and alkylated with iodoacetamide at 8.3 mM. For digestion with trypsin, protein extracts were diluted in 20 mM HEPES pH 8.0 to a final concentration of 2 M urea and soluble TLCK-trypsin (Worthington) was added at 10-20 μg/mL. Digestion was performed for 1-2 days at room temperature. 
     Trifluoroacetic acid (TFA) was added to protein digests to a final concentration of 1 %, precipitate was removed by centrifugation, and digests were loaded onto Sep-Pak C 18  columns (Waters) equilibrated with 0.1% TFA. A column volume of 0.7-1.0 ml was used per 2×10 8  cells. Columns were washed with 15 volumes of 0.1 % TFA, followed by 4 volumes of 5% acetonitrile (MeCN) in 0.1% TFA. Peptide fraction I was obtained by eluting columns with 2 volumes each of 8, 12, and 15% MeCN in 0.1% TFA and combining the eluates. Fractions II and III were a combination of eluates after eluting columns with 18, 22, 25% MeCN in 0.1 % TFA and with 30, 35, 40% MeCN in 0.1% TFA, respectively. All peptide fractions were lyophilized. 
     Peptides from each fraction corresponding to 2×10 8  cells were dissolved in 1 ml of IAP buffer (20 mM Tris/HCl or 50 mM MOPS pH 7.2, 10 mM sodium phosphate, 50 mM NaCl) and insoluble matter (mainly in peptide fractions III) was removed by centrifugation. IAP was performed on each peptide fraction separately. The phosphotyrosine monoclonal antibody P-Tyr-100 (Cell Signaling Technology, Inc., catalog number 9411) was coupled at 4 mg/ml beads to protein G or protein A agarose (Roche), respectively. Immobilized antibody (15 μl, 60 μg) was added as 1:1 slurry in IAP buffer to 1 ml of each peptide fraction, and the mixture was incubated overnight at 4° C. with gentle rotation. The immobilized antibody beads were washed three times with 1 ml IAP buffer and twice with 1 ml water, all at 4° C. Peptides were eluted from beads by incubation with 75 μl of 0.1% TFA at room temperature for 10 minutes. 
     Alternatively, one single peptide fraction was obtained from Sep-Pak C18 columns by elution with 2 volumes each of 10%, 15%, 20%, 25%, 30%, 35% and 40% acetonitirile in 0.1% TFA and combination of all eluates. IAP on this peptide fraction was performed as follows: After lyophilization, peptide was dissolved in 1.4 ml IAP buffer (MOPS pH 7.2, 10 mM sodium phosphate, 50 mM NaCl) and insoluble matter was removed by centrifugation. Immobilized antibody (40 μl, 160 μg) was added as 1:1 slurry in IAP buffer, and the mixture was incubated overnight at 4° C. with gentle shaking. The immobilized antibody beads were washed three times with 1 ml IAP buffer and twice with 1 ml water, all at 4° C. Peptides were eluted from beads by incubation with 55 μl of 0.15% TFA at room temperature for 10 min (eluate 1), followed by a wash of the beads (eluate 2) with 45 μl of 0.15% TFA. Both eluates were combined. 
     Analysis bv LC-MS/MS Mass Spectrometry 
     40 μl or more of IAP eluate were purified by 0.2 μl StageTips or ZipTips. Peptides were eluted from the microcolumns with 1 μl of 40% MeCN, 0.1% TFA (fractions I and II) or 1 μl of 60% MeCN, 0.1% TFA (fraction III) into 7.6 μl of 0.4% acetic acid/0.005% heptafluorobutyric acid. This sample was loaded onto a 10 cm×75 μm PicoFrit capillary column (New Objective) packed with Magic C18 AQ reversed-phase resin (Michrom Bioresources) using a Famos autosampler with an inert sample injection valve (Dionex). The column was then developed with a 45-min linear gradient of acetonitrile delivered at 200 nl/min (Ultimate, Dionex), and tandem mass spectra were collected in a data-dependent manner with an LTQ ion trap mass spectrometer essentially as described by Gygi et al., supra. 
     Database Analysis &amp; Assignments. 
     MS/MS spectra were evaluated using TurboSequest in the Sequest Browser package (v. 27, rev. 12) supplied as part of BioWorks 3.0 (ThermoFinnigan). Individual MS/MS spectra were extracted from the raw data file using the Sequest Browser program CreateDta, with the following settings: bottom MW, 700; top MW, 4,500; minimum number of ions, 20; minimum TIC, 4×10 5 ; and precursor charge state, unspecified. Spectra were extracted from the beginning of the raw data file before sample injection to the end of the eluting gradient. The lonQuest and VuDta programs were not used to further select MS/MS spectra for Sequest analysis. MS/MS spectra were evaluated with the following TurboSequest parameters: peptide mass tolerance, 2.5; fragment ion tolerance, 0.0; maximum number of differential amino acids per modification, 4; mass type parent, average; mass type fragment, average; 
     maximum number of internal cleavage sites, 10; neutral losses of water and ammonia from b and y ions were considered in the correlation analysis. Proteolytic enzyme was specified except for spectra collected from elastase digests. 
     Searches were performed against the NCBI human protein database (as released on Feb. 23, 2004 and containing 27, 175 protein sequences). Cysteine carboxamidomethylation was specified as a static modification, and phosphorylation was allowed as a variable modification on serine, threonine, and tyrosine residues or on tyrosine residues alone. It was determined that restricted phosphorylation to tyrosine residues had little effect on the number of phosphorylation sites assigned. Furthermore it should be noted that certain peptides were originally isolated in mouse and later normalized to human sequences as shown by Tablel/ FIG. 2 . 
     In proteomics research, it is desirable to validate protein identifications based solely on the observation of a single peptide in one experimental result, in order to indicate that the protein is, in fact, present in a sample. This has led to the development of statistical methods for validating peptide assignments, which are not yet universally accepted, and guidelines for the publication of protein and peptide identification results (see Carr et al.,  Mol. Cell Proteomics  3: 531-533 (2004)), which were followed in this Example. However, because the immunoaffinity strategy separates phosphorylated peptides from unphosphorylated peptides, observing just one phosphopeptide from a protein is a common result, since many phosphorylated proteins have only one tyrosine-phosphorylated site. For this reason, it is appropriate to use additional criteria to validate phosphopeptide assignments. Assignments are likely to be correct if any of these additional criteria are met: (i) the same sequence is assigned to co-eluting ions with different charge states, since the MS/MS spectrum changes markedly with charge state; (ii) the site is found in more than one peptide sequence context due to sequence overlaps from incomplete proteolysis or use of proteases other than trypsin; (iii) the site is found in more than one peptide sequence context due to homologous but not identical protein isoforms; (iv) the site is found in more than one peptide sequence context due to homologous but not identical proteins among species; and (v) sites validated by MS/MS analysis of synthetic phosphopeptides corresponding to assigned sequences, since the ion trap mass spectrometer produces highly reproducible MS/MS spectra. The last criterion is routinely employed to confirm novel site assignments of particular interest. 
     All spectra and all sequence assignments made by Sequest were imported into a relational database. The following Sequest scoring thresholds were used to select phosphopeptide assignments that are likely to be correct: RSp&lt;6, XCorr≧2.2, and DeltaCN&gt;0.099. Further, the assigned sequences could be accepted or rejected with respect to accuracy by using the following conservative, two-step process. 
     In the first step, a subset of high-scoring sequence assignments should be selected by filtering for XCorr values of at least 1.5 for a charge state of +1, 2.2 for +2, and 3.3 for +3, allowing a maximum RSp value of 10. Assignments in this subset should be rejected if any of the following criteria were satisfied: (i) the spectrum contains at least one major peak (at least 10% as intense as the most intense ion in the spectrum) that can not be mapped to the assigned sequence as an a, b, or y ion, as an ion arising from neutral-loss of water or ammonia from a b or y ion, or as a multiply protonated ion; (ii) the spectrum does not contain a series of b or y ions equivalent to at least six uninterrupted residues; or (iii) the sequence is not observed at least five times in all the studies conducted (except for overlapping sequences due to incomplete proteolysis or use of proteases other than trypsin). 
     In the second step, assignments with below-threshold scores should be accepted if the low-scoring spectrum shows a high degree of similarity to a high-scoring spectrum collected in another study, which simulates a true reference library-searching strategy. 
     EXAMPLE 2  
     Production of Phospho-Specific Polyclonal Antibodies for the Detection of Leukemia-related Signaling Protein Phosphorylation 
     Polyclonal antibodies that specifically bind a Leukemia-related signal transduction protein only when phosphorylated at the respective phosphorylation site disclosed herein (see Table 1/ FIG. 2 ) are produced according to standard methods by first constructing a synthetic peptide antigen comprising the phosphorylation site sequence and then immunizing an animal to raise antibodies against the antigen, as further described below. Production of exemplary polyclonal antibodies is provided below. 
     A. MELK (tyrosine 438). 
     A 15 amino acid phospho-peptide antigen, SAVKNEEy*FMFPEPK (where y*=phosphotyrosine) that corresponds to the sequence encompassing the tyrosine 438 phosphorylation site in human MELK kinase (see Row 244 of Table 1; SEQ ID NO: 243), plus cysteine on the C-terminal for coupling, is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer. See A NTIBODIES:  A L ABORATORY  M ANUAL,  supra.; Merrifield, supra. This peptide is then coupled to KLH and used to immunize animals to produce (and subsequently screen) phospho-specific MELK(tyr438 ) polyclonal antibodies as described in Immunization/ Screening below. 
     B. SIT1 (tyrosine 127). 
     A 15 amino acid phospho-peptide antigen, AEEVMCy*TSLQLRP (where y*=phosphotyrosine) that corresponds to the sequence encompassing the tyrosine 127 phosphorylation site in human SIT1 adaptor/scaffold protein (see Row 38 of Table 1 (SEQ ID NO: 37)), plus cysteine on the C-terminal for coupling, is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer. See A NTIBODIES:  A L ABORATORY  M ANUAL,  supra.; Merrifield, supra. This peptide is then coupled to KLH and used to immunize animals to produce (and subsequently screen) phospho-specific SIT1(tyr127) polyclonal antibodies as described in Immunization/Screening below. 
     C. PECAM1 (tyrosine 663). 
     A 13 amino acid phospho-peptide antigen, MEANSHy*GHNDDV (where y*=phosphotyrosine) that corresponds to the sequence encompassing the tyrosine 663 phosphorylation site in human PECAM1 adhesion protein (see Row 73 of Table 1 (SEQ ID NO: 72), plus cysteine on the C-terminal for coupling, is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer. See A NTIBODIES:  A L ABORATORY  M ANUAL,  supra.; Merrifield, supra. This peptide is then coupled to KLH and used to immunize animals to produce (and subsequently screen) phospho-specific PECAM1 (tyr663) antibodies as described in Immunization/Screening below. 
     Immunization/Screening. 
     A synthetic phospho-peptide antigen as described in A-C above is coupled to KLH, and rabbits are injected intradermally (ID) on the back with antigen in complete Freunds adjuvant (500 μg antigen per rabbit). The rabbits are boosted with same antigen in incomplete Freund adjuvant (250 μg antigen per rabbit) every three weeks. After the fifth boost, bleeds are collected. The sera are purified by Protein A-affinity chromatography by standard methods (see A NTIBODIES:  A L ABORATORY  M ANUAL,  Cold Spring Harbor, supra.). The eluted immunoglobulins are further loaded onto a non-phosphorylated synthetic peptide antigen-resin Knotes column to pull out antibodies that bind the non-phosphorylated form of the phosphorylation site. The flow through fraction is collected and applied onto a phospho-synthetic peptide antigen-resin column to isolate antibodies that bind the phosphorylated form of the site. After washing the column extensively, the bound antibodies (i.e. antibodies that bind a phosphorylated peptide described in A-C above, but do not bind the non-phosphorylated form of the peptide) are eluted and kept in antibody storage buffer. 
     The isolated antibody is then tested for phospho-specificity using Western blot assay using an appropriate cell line that expresses (or overexpresses) target phospho-protein (i.e. phosphorylated MELK, SIT1 or PECAM1), for example, SEM, Jurkat and MKPL-1 cells, respectively. Cells are cultured in DMEM or RPMI supplemented with 10% FCS. Cell are collected, washed with PBS and directly lysed in cell lysis buffer. The protein concentration of cell lysates is then measured. The loading buffer is added into cell lysate and the mixture is boiled at 100° C. for 5 minutes. 20 μl (10 μg protein) of sample is then added onto 7.5% SDS-PAGE gel. 
     A standard Western blot may be performed according to the Immunoblotting Protocol set out in the C ELL  S IGNALING  T ECHNOLOGY,  I NC.  2003-04 Catalogue, p. 390. The isolated phospho-specific antibody is used at dilution 1:1000. Phosphorylation-site specificity of the antibody will be shown by binding of only the phosphorylated form of the target protein. Isolated phospho-specific polyclonal antibody does not (substantially) recognize the target protein when not phosphorylated at the appropriate phosphorylation site in the non-stimulated cells (e.g. PECAM1 is not bound when not phosphorylated at tyrosine 663). 
     In order to confirm the specificity of the isolated antibody, different cell lysates containing various phosphorylated signal transduction proteins other than the target protein are prepared. The Western blot assay is performed again using these cell lysates. The phospho-specific polyclonal antibody isolated as described above is used (1:1000 dilution) to test reactivity with the different phosphorylated non-target proteins on Western blot membrane. The phospho-specific antibody does not significantly cross-react with other phosphorylated signal transduction proteins, although occasionally slight binding with a highly homologous phosphorylation-site on another protein may be observed. In such case the antibody may be further purified using affinity chromatography, or the specific immunoreactivity cloned by rabbit hybridoma technology. 
     EXAMPLE 3  
     Production of Phospho-Specific Monoclonal Antibodies for the Detection of Leukemia-Related Signaling Protein Phosphorylation 
     Monoclonal antibodies that specifically bind a Leukemia-related signal transduction protein only when phosphorylated at the respective phosphorylation site disclosed herein (see Table 1/ FIG. 2 ) are produced according to standard methods by first constructing a synthetic peptide antigen comprising the phosphorylation site sequence and then immunizing an animal to raise antibodies against the antigen, and harvesting spleen cells from such animals to produce fusion hybridomas, as further described below. Production of exemplary monoclonal antibodies is provided below. 
     A. TSC2 (tyrosine 1736). 
     A 10 amino acid phospho-peptide antigen, PTDIy*PSKW (where y*=phosphotyrosine) that corresponds to the sequence encompassing the tyrosine 1736 phosphorylation site in human TSC2 GTPase activating protein (see Row 87 of Table 1 (SEQ ID NO: 86)), plus cysteine on the C-terminal for coupling, is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer. See A NTIBODIES:  A L ABORATORY  M ANUAL,  supra.; Merrifield, supra. This peptide is then coupled to KLH and used to immunize animals and harvest spleen cells for generation (and subsequent screening) of phospho-specific monoclonal TSC2(tyr1736) antibodies as described in Immunization/Fusion/Screening below. 
     B. CD84 (tyrosine 279). 
     A 10 amino acid phospho-peptide antigen, ESRIy*DEIL (where y*=phosphotyrosine) that corresponds to the sequence encompassing the tyrosine 279 phosphorylation site in human CD84 cell surface protein (see Row 93 of Table 1 (SEQ ID NO: 92)), plus cysteine on the C-terminal for coupling, is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer. See A NTIBODIES:  A L ABORATORY  M ANUAL,  supra.; Merrifield, supra. This peptide is then coupled to KLH and used to immunize animals and harvest spleen cells for generation (and subsequent screening) of phospho-specific monoclonal CD84(tyr279) antibodies as described in Immunization/Fusion/Screening below. 
     C. STAT5A (tyrosine 22). 
     A 14 amino acid phospho-peptide antigen, QMQVLy*GQHFPIEV (where Y*=phosphotyrosine) that corresponds to the sequence encompassing the tyrosine 22 phosphorylation site in human STAT5A transcription factor (see Row 411 of Table 1 (SEQ ID NO: 410)), plus cysteine on the C-terminal for coupling, is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer. See A NTIBODIES:  A L ABORATORY  M ANUAL,  supra.; Merrifield, supra. This peptide is then coupled to KLH and used to immunize animals and harvest spleen cells for generation (and subsequent screening) of phospho-specific monoclonal STAT5A (tyr22) antibodies as described in Immunization/Fusion/Screening below. 
     Immunization/Fusion/Screening. 
     A synthetic phospho-peptide antigen as described in A-C above is coupled to KLH, and BALB/C mice are injected intradermally (ID) on the back with antigen in complete Freunds adjuvant (e.g. 50 μg antigen per mouse). The mice are boosted with same antigen in incomplete Freund adjuvant (e.g. 25 μg antigen per mouse) every three weeks. After the fifth boost, the animals are sacrificed and spleens are harvested. 
     Harvested spleen cells are fused to SP2/0 mouse myeloma fusion partner cells according to the standard protocol of Kohler and Milstein (1975). Colonies originating from the fusion are screened by ELISA for reactivity to the phospho-peptide and non-phospho-peptide forms of the antigen and by Western blot analysis (as described in Example 1 above). Colonies found to be positive by ELISA to the phospho-peptide while negative to the non-phospho-peptide are further characterized by Western blot analysis. Colonies found to be positive by Western blot analysis are subcloned by limited dilution. Mouse ascites are produced from a single clone obtained from subcloning, and tested for phospho-specificity (against the TSC2, CD84, or STAT5A phospho-peptide antigen, as the case may be) on ELISA. Clones identified as positive on Western blot analysis using cell culture supernatant as having phospho-specificity, as indicated by a strong band in the induced lane and a weak band in the uninduced lane of the blot, are isolated and subcloned as clones producing monoclonal antibodies with the desired specificity. 
     Ascites fluid from isolated clones may be further tested by Western blot analysis. The ascites fluid should produce similar results on Western blot analysis as observed previously with the cell culture supernatant, indicating phospho-specificity against the phosphorylated target (e.g. STAT5A phosphorylated at tyrosine 22). 
     EXAMPLE 4  
     Production and Use of AQUA Peptides for the Quantification of Leukemia-Related Signaling Protein Phosphorylation 
     Heavy-isotope labeled peptides (AQUA peptides (internal standards)) for the detection and quantification of a Leukemia-related signal transduction protein only when phosphorylated at the respective phosphorylation site disclosed herein (see Table 1/ FIG. 2 ) are produced according to the standard AQUA methodology (see Gygi et al., Gerber et al., supra.) methods by first constructing a synthetic peptide standard corresponding to the phosphorylation site sequence and incorporating a heavy-isotope label. Subsequently, the MS n  and LC-SRM signature of the peptide standard is validated, and the AQUA peptide is used to quantify native peptide in a biological sample, such as a digested cell extract. Production and use of exemplary AQUA peptides is provided below. 
     A. ZAP70 (tyrosine 164). 
     An AQUA peptide comprising the sequence, MPWy*HSSLTR (y*=phosphotyrosine; sequence incorporating  14 C/ 15 N-labeled leucine (indicated by bold L), which corresponds to the tyrosine 164 phosphorylation site in human ZAP70 kinase (see Row 284 in Table 1 (SEQ ID NO: 283)), is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer (see Merrifield, supra.) as further described below in Synthesis &amp; MS/MS Signature. The ZAP70(tyr164) AQUA peptide is then spiked into a biological sample to quantify the amount of phosphorylated ZAP70(tyr164) in the sample, as further described below in Analysis &amp; Quantification. 
     B. SCAP1 (tyrosine 142). 
     An AQUA peptide comprising the sequence GLFYy*YANEK (y*=phosphotyrosine; sequence incorporating  14 C/ 15 N-labeled leucine (indicated by bold L), which corresponds to the tyrosine 142 phosphorylation site in human SCAP1 adaptor/scaffold protein (see Row 42 in Table 1 (SEQ ID NO: 41)), is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer (see Merrifield, supra.) as further described below in Synthesis &amp; MS/MS Signature. The SCAP1(tyr142) AQUA peptide is then spiked into a biological sample to quantify the amount of phosphorylated SCAP1(tyr142) in the sample, as further described below in Analysis &amp; Quantification. 
     C. CFL1 (tyrosine 68) 
     An AQUA peptide comprising the sequence, GQTVDDPy*ATFVKML (y*=phosphotyrosine; sequence incorporating  14 C/ 15 N-labeled phenylalanine (indicated by bold F), which corresponds to the tyrosine 211 phosphorylation site in human CFL1 adaptor/scaffold protein (see Row 121 in Table 1 (SEQ ID NO: 120)), is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer (see Merrifield, supra.) as further described below in Synthesis &amp; MS/MS Signature. The CFL1(tyr68) AQUA peptide is then spiked into a biological sample to quantify the amount of phosphorylated CFL1 (tyr68) in the sample, as further described below in Analysis &amp; Quantification. 
     D. BLK (tyrosine 187). 
     An AQUA peptide comprising the sequence, CLDEGGy*YISPR (y*=phosphotyrosine; sequence incorporating  14 C/ 15 N-labeled proline (indicated by bold P), which corresponds to the tyrosine 187 phosphorylation site in human BLK kinase (see Row 271 in Table 1 (SEQ ID NO: 270)), is constructed according to standard synthesis techniques using, e.g., a Rainin/Protein Technologies, Inc., Symphony peptide synthesizer (see Merrifield, supra.) as further described below in Synthesis &amp; MS/MS Signature. The BLK(tyr187) AQUA peptide is then spiked into a biological sample to quantify the amount of phosphorylated BLK(tyr187) in the sample, as further described below in Analysis &amp; Quantification. 
     Synthesis &amp; MS/MS Spectra. 
     Fluorenylmethoxycarbonyl (Fmoc)-derivatized amino acid monomers may be obtained from AnaSpec (San Jose, Calif.). Fmoc-derivatized stable-isotope monomers containing one  15 N and five to nine  13 C atoms may be obtained from Cambridge Isotope Laboratories (Andover, Mass.). Preloaded Wang resins may be obtained from Applied Biosystems. Synthesis scales may vary from 5 to 25 μmol. Amino acids are activated in situ with 1-H-benzotriazolium, 1-bis(dimethylamino)methylene]-hexafluorophosphate (1-),3-oxide:1-hydroxybenzotriazole hydrate and coupled at a 5-fold molar excess over peptide. Each coupling cycle is followed by capping with acetic anhydride to avoid accumulation of one-residue deletion peptide by-products. After synthesis peptide-resins are treated with a standard scavenger-containing trifluoroacetic acid (TFA)-water cleavage solution, and the peptides are precipitated by addition to cold ether. Peptides (i.e. a desired AQUA peptide described in A-D above) are purified by reversed-phase C18 HPLC using standard TFA/acetonitrile gradients and characterized by matrix-assisted laser desorption ionization-time of flight (Biflex III, Bruker Daltonics, Billerica, Mass.) and ion-trap (ThermoFinnigan, LCQ DecaXP) MS. 
     MS/MS spectra for each AQUA peptide should exhibit a strong y-type ion peak as the most intense fragment ion that is suitable for use in an SRM monitoring/analysis. Reverse-phase microcapillary columns (0.1 Å˜150-220 mm) are prepared according to standard methods. An Agilent 1100 liquid chromatograph may be used to develop and deliver a solvent gradient [0.4% acetic acid/0.005% heptafluorobutyric acid (HFBA)/7% methanol and 0.4% acetic acid/0.005% HFBA/65% methanol/35% acetonitrile] to the microcapillary column by means of a flow splitter. Samples are then directly loaded onto the microcapillary column by using a FAMOS inert capillary autosampler (LC Packings, San Francisco) after the flow split. Peptides are reconstituted in 6% acetic acid/0.01% TFA before injection. 
     Analysis &amp; Quantification. 
     Target protein (e.g. a phosphorylated protein of A-D above) in a biological sample is quantified using a validated AQUA peptide (as described above). The IAP method is then applied to the complex mixture of peptides derived from proteolytic cleavage of crude cell extracts to which the AQUA peptides have been spiked in. 
     LC-SRM of the entire sample is then carried out. MS/MS may be performed by using a ThermoFinnigan (San Jose, Calif.) mass spectrometer (LTQ ion trap or TSQ Quantum triple quadrupole). On the LTQ, parent ions are isolated at 1.6 m/z width, the ion injection time being limited to 100 ms per microscan, with one microscans per peptide, and with an AGC setting of 1×10 5 ; on the Quantum, Q1 is kept at 0.4 and Q3 at 0.8 m/z with a scan time of 200 ms per peptide. On both instruments, analyte and internal standard are analyzed in alternation within a previously known reverse-phase retention window; well-resolved pairs of internal standard and analyte are analyzed in separate retention segments to improve duty cycle. Data are processed by integrating the appropriate peaks in an extracted ion chromatogram (60.15 m/z from the fragment monitored) for the native and internal standard, followed by calculation of the ratio of peak areas multiplied by the absolute amount of internal standard (e.g., 500 fmol).