Abstract:
Long QT Syndrome (LQTS) is a cardiovascular disorder characterized by prolongation of the QT interval on electrocardiogram and presence of syncope, seizures and sudden death. Five genes have been implicated in Romano-Ward syndrome, the autosomal dominant form of LQTS. These genes are KVLQT1, HERG, SCN5A, KCNE1 and KCNE2. Mutations in KVLQt1 and KCNE1 also cause the Jervell and Lange-Nielsen syndrome, a form of LQTS associated with deafness, a phenotypic abnormality inherited in an autosomal recessive fashion. Mutational analyses were used to screen 262 unrelated individuals with LQTS for mutations in the five defined genes. A total of 134 mutations were observed of which eighty were novel.

Description:
CROSS REFERENCE TO RELATED APPLICATIONS  
       [0001]    The present invention is related to provisional application Ser. No. 60/190,057 filed Mar. 17, 2000, and is also related to provisional application Ser. No. 60/147,488 filed Aug. 9, 1999, both of which are incorporated herein by reference. 
     
    
       [0002] This application was made with Government support from NHLBI under Grant Nos. RO1-HL46401, RO1-HL33843, RO1-HL51618, P50-HL52338 and MO1-RR000064. The federal government may have certain rights in this invention. 
     
    
     
       BACKGROUND OF THE INVENTION  
         [0003]    Long QT Syndrome (LQTS) is a cardiovascular disorder characterized by prolongation of the QT interval on electrocardiogram and presence of syncope, seizures and sudden death, usually in young, otherwise healthy individuals (Jervell and Lange-Nielsen, 1957; Romano et al., 1963; Ward, 1964). The clinical features of LQTS result from episodic ventricular tachyarrhythmias, such as torsade de pointes and ventricular fibrillation (Schwartz et al., 1975; Moss et al., 1991). Two inherited forms of LQTS exist. The more common form, Romano-Ward syndrome (RW), is not associated with other phenotypic abnormalities and is inherited as an autosomal dominant trait with variable penetrance (Roman et al., 1963; Ward, 1964). Jervell and Lange-Nielsen syndrome (JLN) is characterized by the presence of deafness, a phenotypic abnormality inherited as an autosomal recessive trait (Jervell and Lange-Nielsen, 1957). LQTS can also be acquired, usually as a result of pharmacologic therapy.  
           [0004]    In previous studies, we mapped LQTS loci to chromosomes 11p15.5 (LQT1) (Keating et al., 1991), 7 q35-36 (LQT2) (Jiang et al., 1994) and LQT3 to 3p21-24 (Jiang et al., 1994). A fourth locus (LQT4) was mapped to 4q25-27 (Schott et al., 1995). Five genes have been implicated in Romano-Ward syndrome, the autosomal dominant form of LQTS. These genes are KVLQT1 (LQT1) (Wang Q. et al., 1996a), HERG (LQT2) (Curran et al., 1995), SCN5A (LQT3) (Wang et al., 1995a), and two genes located at 21q22-KCNE1 (LQT5) (Splawski et al., 1997a) and KCNE2 (LQT6) (Abbott et al., 1999). Mutations in KVLQT1 and KCNE1 also cause the Jervell and Lange-Nielsen syndrome, a form of LQTS associated with deafness, a phenotypic abnormality inherited in an autosomal recessive fashion.  
           [0005]    KVLQT1, HERG, KCNE1 and KCNE2 encode potassium channel subunits. Four KVLQT1 α-subunits assemble with minK (β-subunits encoded by KCNE1, stoichiometry is unknown) to form I Ks  channels underlying the slowly activating delayed rectifier potassium current in the heart (Sanguinetti et al., 1996a; Barhanin et al., 1996). Four HERG α-subunits assemble with MiRP1 (encoded by KCNE2, stoichiometry unknown) to form I Kr  channels, which underlie the rapidly activating, delayed rectifier potassium current (Abbott et al., 1999). Mutant subunits lead to reduction of I Ks  or I Kr  by a loss-of-function mechanism, often with a dominant-negative effect (Chouabe et al., 1997; Shalaby et al., 1997; Wollnik et al., 1997; Sanguinetti et al., 1996b). SCN5A encodes the cardiac sodium channel that is responsible for I Na , the sodium current in the heart (Gellens et al., 1992). LQTS-associated mutations in SCN5A cause a gain-of-function (Bennett et al., 1995; Dumaine et al., 1996). In the heart, reduced I Ks  or I Kr  or increased I Na  leads to prolongation of the cardiac action potential, lengthening of the QT interval and increased risk of arrhythmia. KVLQT1 and KCNE1 are also expressed in the inner ear (Neyroud et al., 1997; Vetter et al., 1996). Others and we demonstrated that complete loss of I Ks  causes the severe cardiac phenotype and deafness in JLN (Neyroud et al., 1997; Splawski et al., 1997b; Tyson et al., 1997; Schulze-Bahr et al., 1997).  
           [0006]    Presymptomatic diagnosis of LQTS is currently based on prolongation of the QT interval on electrocardiogram. Genetic studies, however, have shown that diagnosis based solely on electrocardiogram is neither sensitive nor specific (Vincent et al., 1992; Priori et al., 1999). Genetic screening using mutational analysis can improve presymptomatic diagnosis. However, a comprehensive study identifying and cataloging all LQTS-associated mutations in all five genes has not been achieved. To determine the relative frequency of mutations in each gene, facilitate presymptomatic diagnosis and enable genotype-phenotype studies, we screened a pool of 262 unrelated individuals with LQTS for mutations in the five defined genes. The results of these studies are presented in the Examples below.  
           [0007]    The present invention relates to alterations in the KVLQT1, HERG, SCN5A, KCNE1 and KCNE2 genes and methods for detecting such alterations.  
           [0008]    The publications and other materials used herein to illuminate the background of the invention or provide additional details respecting the practice, are incorporated by reference, and for convenience are respectively grouped in the appended List of References.  
           [0009]    The present invention is directed to alterations in genes and gene products associated with long QT syndrome and to a process for the diagnosis and prevention of LQTS. LQTS diagnosed in accordance with the present invention by analyzing the DNA sequence of the KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 gene of an individual to be tested and comparing the respective DNA sequence to the known DNA sequence of the normal gene. Alternatively, these genes of an individual to be tested can be screened for mutations which cause LQTS. Prediction of LQTS will enable practitioners to prevent this disorder using existing medical therapy.  
         SUMMARY OF THE INVENTION  
         [0010]    The present invention relates to alterations in the KVLQT1, HERG, SCN5A, KCNE1 and KCNE2 genes and methods for detecting such alterations. The alterations in the KVLQT1, HERG, SCN5A, KCNE1 and KCNE2 genes include mutations and polymorphisms. Included among the mutations are frameshift, nonsense, splice, regulatory and missense mutations. Any method which is capable of detecting the alterations described herein can be used. Such methods include, but are not limited to, DNA sequencing, allele-specific probing, mismatch detection, single stranded conformation polymorphism detection and allele-specific PCR amplification. 
       
    
    
     BRIEF DESCRIPTION OF THE FIGURES  
       [0011]    [0011]FIG. 1 is a schematic representation of the predicted topology of KVLQT1 and the locations of LQTS-associated mutations. KVLQT1 consists of six putative transmembrane segments (S1 to S6) and a pore (Pore) region. Each circle represents an amino acid. The approximate location of LQTS-associated mutations identified in our laboratory are shown with filled circles.  
         [0012]    [0012]FIG. 2 is a schematic representation of HERG mutations. HERG consists of six putative transmembrane segments (S1 to S6) and a pore (Pore) region. Location of LQTS-associated mutations are shown with filled circles.  
         [0013]    [0013]FIG. 3 is a schematic representation of SCN5A and locations of LQTS-associated mutations. SCN5A consists of four domain (DI to DIV), each of which has six putative transmembrane segments (S1 to S6) and a pore (Pore) region. Location of LQTS-associated mutations identified in our laboratory are shown with filled circles.  
         [0014]    [0014]FIG. 4 is a schematic representation of minK and locations of LQT-associated mutations. MinK consists of one putative transmembrane domain (SI). The approximate location of LQTS-associated mutations identified in our laboratory are shown with filled circles.  
         [0015]    [0015]FIG. 5 is a schematic representation of the predicted topology of MiRP1 and locations of arrhythmia-associated mutations. MiRPI consists of one putative transmembrane domain (S1). The approximate location of arrhythmia-associated mutations identified in our laboratory are shown with filled circles. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0016]    The present invention relates to alterations in the KVLQT1, HERG, SCN5A, KCNE1 and KCNE2 genes and methods for detecting such alterations. The alterations in the KVLQT1, HERG, SCN5A, KCNE1 and KCNE2 genes include mutations and polymorphisms. Included among the mutations are frameshift, nonsense, splice, regulatory and missense mutations. Any method which is capable of detecting the mutations and polymorphisms described herein can be used. Such methods include, but are not limited to, DNA sequencing, allele-specific probing, mismatch detection, single stranded conformation polymorphism detection and allele-specific PCR amplification.  
         [0017]    KVLQT1, HERG, SCN5A, KCNE1 and KCNE2 mutations cause increased risk for LQTS. Many different mutations occur in KVLQT1, HERG, SCN5A, KCNE1 and KCNE2. In order to detect the presence of alterations in the KVLQT1, HERG, SCN5A, KCNE1 and KCNE2 genes, a biological sample such as blood is prepared and analyzed for the presence or absence of a given alteration of KVLQT1, HERG, SCN5A, KCNE1 or KCNE2. In order to detect the increased risk for LQTS or for the lack of such increased risk, a biological sample is prepared and analyzed for the presence or absence of a mutant allele of KVLQT1, HERG, SCN5A, KCNE1 or KCNE2. Results of these tests and interpretive information are returned to the health care provider for communication to the tested individual. Such diagnoses may be performed by diagnostic laboratories or, alternatively, diagnostic kits are manufactured and sold to health care providers or to private individuals for self-diagnosis.  
         [0018]    The presence of hereditary LQTS may be ascertained by testing any tissue of a human for mutations of the KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 gene. For example, a person who has inherited a germline HERG mutation would be prone to develop LQTS. This can be determined by testing DNA from any tissue of the person&#39;s body. Most simply, blood can be drawn and DNA extracted from the cells of the blood. In addition, prenatal diagnosis can be accomplished by testing fetal cells, placental cells or amniotic cells for mutations of the KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 gene. Alteration of a wild-type KVLQT1, HERG, SCNSA, KCNEI or KCNE2 allele, whether, for example, by point mutation or deletion, can be detected by any of the means discussed herein.  
         [0019]    There are several methods that can be used to detect DNA sequence variation. Direct DNA sequencing, either manual sequencing or automated fluorescent sequencing can detect sequence variation. Another approach is the single-stranded conformation polymorphism assay (SSCP) (Orita et al., 1989). This method does not detect all sequence changes, especially if the DNA fragment size is greater than 200 bp, but can be optimized to detect most DNA sequence variation. The reduced detection sensitivity is a disadvantage, but the increased throughput possible with SSCP makes it an attractive, viable alternative to direct sequencing for mutation detection on a research basis. The fragments which have shifted mobility on SSCP gels are then sequenced to determine the exact nature of the DNA sequence variation. Other approaches based on the detection of mismatches between the two complementary DNA strands include clamped denaturing gel electrophoresis (CDGE) (Sheffield et al., 1991), heteroduplex analysis (HA) (White et al., 1992) and chemical mismatch cleavage (CMC) (Grompe et al., 1989). None of the methods described above will detect large deletions, duplications or insertions, nor will they detect a regulatory mutation which affects transcription or translation of the protein. Other methods which might detect these classes of mutations such as a protein truncation assay or the asymmetric assay, detect only specific types of mutations and would not detect missense mutations. A review of currently available methods of detecting DNA sequence variation can be found in a recent review by Grompe (1993). Once a mutation is known, an allele specific detection approach such as allele specific oligonucleotide (ASO) hybridization can be utilized to rapidly screen large numbers of other samples for that same mutation. Such a technique can utilize probes which are labeled with gold nanoparticles to yield a visual color result (Elghanian et al., 1997).  
         [0020]    A rapid preliminary analysis to detect polymorphisms in DNA sequences can be performed by looking at a series of Southern blots of DNA cut with one or more restriction enzymes, preferably with a large number of restriction enzymes. Each blot contains a series of normal individuals and a series of LQTS cases. Southern blots displaying hybridizing fragments (differing in length from control DNA when probed with sequences near or including the HERG locus) indicate a possible mutation. If restriction enzymes which produce very large restriction fragments are used, then pulsed field gel electrophoresis (PFGE) is employed.  
         [0021]    Detection of point mutations may be accomplished by molecular cloning of the KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 alleles and sequencing the alleles using techniques well known in the art. Also, the gene or portions of the gene may be amplified, e.g., by PCR or other amplification technique, and the amplified gene or amplified portions of the gene may be sequenced.  
         [0022]    There are six well known methods for a more complete, yet still indirect, test for confirming the presence of a susceptibility allele: 1) single stranded conformation analysis (SSCP) (Orita et al., 1989); 2) denaturing gradient gel electrophoresis (DGGE) (Wartell et al., 1990; Sheffield et al., 1989); 3) RNase protection assays (Finkelstein et al., 1990; Kinszler et al., 1991); 4) allele-specific oligonucleotides (ASOs) (Conner et al., 1983); 5) the use of proteins which recognize nucleotide mismatches, such as the  E. coli  mutS protein (Modrich, 1991); and 6) allele-specific PCR (Ruano and Kidd, 1989). For allele-specific PCR, primers are used which hybridize at their 3′ ends to a particular KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 mutation. If the particular mutation is not present, an amplification product is not observed. Amplification Refractory Mutation System (ARMS) can also be used, as disclosed in European Patent Application Publication No. 0332435 and in Newton et al., 1989. Insertions and deletions of genes can also be detected by cloning, sequencing and amplification. In addition, restriction fragment length polymorphism (RFLP) probes for the gene or surrounding marker genes can be used to score alteration of an allele or an insertion in a polymorphic fragment. Such a method is particularly useful for screening relatives of an affected individual for the presence of the mutation found in that individual. Other techniques for detecting insertions and deletions as known in the art can be used.  
         [0023]    In the first three methods (SSCP, DGGE and RNase protection assay), a new electrophoretic band appears. SSCP detects a band which migrates differentially because the sequence change causes a difference in single-strand, intramolecular base pairing. RNase protection involves cleavage of the mutant polynucleotide into two or more smaller fragments. DGGE detects differences in migration rates of mutant sequences compared to wild-type sequences, using a denaturing gradient gel. In an allele-specific oligonucleotide assay, an oligonucleotide is designed which detects a specific sequence, and the assay is performed by detecting the presence or absence of a hybridization signal. In the mutS assay, the protein binds only to sequences that contain a nucleotide mismatch in a heteroduplex between mutant and wild-type sequences.  
         [0024]    Mismatches, according to the present invention, are hybridized nucleic acid duplexes in which the two strands are not 100% complementary. Lack of total homology may be due to deletions, insertions, inversions or substitutions. Mismatch detection can be used to detect point mutations in the gene or in its mRNA product. While these techniques are less sensitive than sequencing, they are simpler to perform on a large number of samples. An example of a mismatch cleavage technique is the RNase protection method. In the practice of the present invention, the method involves the use of a labeled riboprobe which is complementary to the human wild-type KVLQT1, HERG, SCN5A, KCNEI or KCNE2 gene coding sequence. The riboprobe and either mRNA or DNA isolated from the person are annealed (hybridized) together and subsequently digested with the enzyme RNase A which is able to detect some mismatches in a duplex RNA structure. If a mismatch is detected by RNase A, it cleaves at the site of the mismatch. Thus, when the annealed RNA preparation is separated on an electrophoretic gel matrix, if a mismatch has been detected and cleaved by RNase A, an RNA product will be seen which is smaller than the full length duplex RNA for the riboprobe and the mRNA or DNA. The riboprobe need not be the full length of the mRNA or gene but can be a segment of either. If the riboprobe comprises only a segment of the mRNA or gene, it will be desirable to use a number of these probes to screen the whole mRNA sequence for mismatches.  
         [0025]    In similar fashion, DNA probes can be used to detect mismatches, through enzymatic or chemical cleavage. See, e.g., Cotton et al., 1988; Shenk et al., 1975; Novack et al., 1986. Alternatively, mismatches can be detected by shifts in the electrophoretic mobility of mismatched duplexes relative to matched duplexes. See, e.g., Cariello, 1988. With either riboprobes or DNA probes, the cellular mRNA or DNA which might contain a mutation can be amplified using PCR (see below) before hybridization. Changes in DNA of the KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 gene can also be detected using Southern hybridization, especially if the changes are gross rearrangements, such as deletions and insertions.  
         [0026]    DNA sequences of the KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 gene which have been amplified by use of PCR may also be screened using allele-specific probes. These probes are nucleic acid oligomers, each of which contains a region of the gene sequence harboring a known mutation. For example, one oligomer may be about 30 nucleotides in length, corresponding to a portion of the gene sequence. By use of a battery of such allele-specific probes, PCR amplification products can be screened to identify the presence of a previously identified mutation in the gene. Hybridization of allele-specific probes with amplified KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 sequences can be performed, for example, on a nylon filter. Hybridization to a particular probe under high stringency hybridization conditions indicates the presence of the same mutation in the tissue as in the allele-specific probe.  
         [0027]    The newly developed technique of nucleic acid analysis via microchip technology is also applicable to the present invention. In this technique, literally thousands of distinct oligonucleotide probes are built up in an array on a silicon chip. Nucleic acid to be analyzed is fluorescently labeled and hybridized to the probes on the chip. It is also possible to study nucleic acid-protein interactions using these nucleic acid microchips. Using this technique one can determine the presence of mutations or even sequence the nucleic acid being analyzed or one can measure expression levels of a gene of interest. The method is one of parallel processing of many, even thousands, of probes at once and can tremendously increase the rate of analysis. Several papers have been published which use this technique. Some of these are Hacia et al., 1996; Shoemaker et al., 1996; Chee et al., 1996; Lockhart et al., 1996; DeRisi et al., 1996; Lipshutz et al., 1995. This method has already been used to screen people for mutations in the breast cancer gene BRCA1 (Hacia et al., 1996). This new technology has been reviewed in a news article in Chemical and Engineering News (Borman, 1996) and been the subject of an editorial (Editorial, Nature Genetics, 1996). Also see Fodor (1997).  
         [0028]    The most definitive test for mutations in a candidate locus is to directly compare genomic KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 sequences from patients with those from a control population. Alternatively, one could sequence messenger RNA after amplification, e.g., by PCR, thereby eliminating the necessity of determining the exon structure of the candidate gene.  
         [0029]    Mutations from patients falling outside the coding region of KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 can be detected by examining the non-coding regions, such as introns and regulatory sequences near or within the genes An early indication that mutations in noncoding regions are important may come from Northern blot experiments that reveal messenger RNA molecules of abnormal size or abundance in patients as compared to control individuals.  
         [0030]    Alteration of KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 mRNA expression can be detected by any techniques known in the art. These include Northern blot analysis, PCR amplification and RNase protection. Diminished mRNA expression indicates an alteration of the wild-type gene. Alteration of wild-type genes can also be detected by screening for alteration of wild-type KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 protein. For example, monoclonal antibodies immunoreactive with HERG can be used to screen a tissue. Lack of cognate antigen would indicate a mutation. Antibodies specific for products of mutant alleles could also be used to detect mutant gene product. Such immunological assays can be done in any convenient formats known in the art. These include Western blots, immunohistochemical assays and ELISA assays. Any means for detecting an altered KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 protein can be used to detect alteration of wild-type KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 genes. Functional assays, such as protein binding determinations, can be used. In addition, assays can be used which detect KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 biochemical function. Finding a mutant KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 gene product indicates alteration of a wild-type KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 gene.  
         [0031]    Mutant KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 genes or gene products can also be detected in other human body samples, such as serum, stool, urine and sputum. The same techniques discussed above for detection of mutant genes or gene products in tissues can be applied to other body samples. By screening such body samples, a simple early diagnosis can be achieved for hereditary LQTS.  
         [0032]    Initially, the screening method involves amplification of the relevant KVLQT1, HERG, SCN5A, KCNE1 or KCNE2 sequence. In another preferred embodiment of the invention, the screening method involves a non-PCR based strategy. Such screening methods include two-step label amplification methodologies that are well known in the art. Both PCR and non-PCR based screening strategies can detect target sequences with a high level of sensitivity. Further details of these methods are briefly presented below and further descriptions can be found in PCT published application WO 96/05306, incorporated herein by reference.  
         [0033]    The most popular method used today is target amplification. Here, the target nucleic acid sequence is amplified with polymerases. One particularly preferred method using polymerase-driven amplification is the polymerase chain reaction (PCR). The polymerase chain reaction and other polymerase-driven amplification assays can achieve over a million-fold increase in copy number through the use of polymerase-driven amplification cycles. Once amplified, the resulting nucleic acid can be sequenced or used as a substrate for DNA probes.  
         [0034]    When the probes are used to detect the presence of the target sequences, the biological sample to be analyzed, such as blood or serum, may be treated, if desired, to extract the nucleic acids. The sample nucleic acid may be prepared in various ways to facilitate detection of the target sequence; e.g. denaturation, restriction digestion, electrophoresis or dot blotting. The targeted region of the analyte nucleic acid usually must be at least partially single-stranded to form hybrids with the targeting sequence of the probe. If the sequence is naturally single-stranded, denaturation will not be required. However, if the sequence is double-stranded, the sequence will probably need to be denatured. Denaturation can be carried out by various techniques known in the art.  
         [0035]    Analyte nucleic acid and probe are incubated under conditions which promote stable hybrid formation of the target sequence in the probe with the putative targeted sequence in the analyte. The region of the probes which is used to bind to the analyte can be made completely complementary to the targeted region of the genes. Therefore, high stringency conditions are desirable in order to prevent false positives. However, conditions of high stringency are used only if the probes are complementary to regions of the chromosome which are unique in the genome. The stringency of hybridization is determined by a number of factors during hybridization and during the washing procedure, including temperature, ionic strength, base composition, probe length, and concentration of formamide. Under certain circumstances, the formation of higher order hybrids, such as triplexes, quadraplexes, etc., may be desired to provide the means of detecting target sequences.  
         [0036]    Detection, if any, of the resulting hybrid is usually accomplished by the use of labeled probes. Alternatively, the probe may be unlabeled, but may be detectable by specific binding with a ligand which is labeled, either directly or indirectly. Suitable labels, and methods for labeling probes and ligands are known in the art, and include, for example, radioactive labels which may be incorporated by known methods (e.g., nick translation, random priming or kinasing), biotin, fluorescent groups, chemiluminescent groups (e.g., dioxetanes, particularly triggered dioxetanes), enzymes, antibodies nd the like. Variations of this basic scheme known in the art, and include those variations that facilitate separation of the hybrids to be detected from extraneous materials and/or that amplify the signal from the labeled moiety. A number of these variations are well known.  
         [0037]    As noted above, non-PCR based screening assays are also contemplated in this invention. This procedure hybridizes a nucleic acid probe (or an analog such as a methyl phosphonate backbone replacing the normal phosphodiester), to the low level DNA target. This probe may have an enzyme covalently linked to the probe, such that the covalent linkage does not interfere with the specificity of the hybridization. This enzyme-probe-conjugate-target nucleic acid complex can then be isolated away from the free probe enzyme conjugate and a substrate is added for enzyme detection. Enzymatic activity is observed as a change in color development or luminescent output resulting in a 10 3 -10 6  increase in sensitivity. For example, the preparation of oligodeoxynucleotide-alkaline phosphatase conjugates and their use as hybridization probes are well known.  
         [0038]    Two-step label amplification methodologies are known in the art. These assays work on the principle that a small ligand (such as digoxigenin, biotin, or the like) is attached to a nucleic acid probe capable of specifically binding the target gene. Allele specific probes are also contemplated within the scope of this example.  
         [0039]    In one example, the small ligand attached to the nucleic acid probe is specifically recognized by an antibody-enzyme conjugate. In one embodiment of this example, digoxigenin is attached to the nucleic acid probe. Hybridization is detected by an antibody-alkaline phosphatase conjugate which turns over a chemiluminescent substrate. In a second example, the small ligand is recognized by a second ligand-enzyme conjugate that is capable of specifically complexing to the first ligand. A well known embodiment of this example is the biotin-avidin type of interactions. Methods for labeling nucleic acid probes and their use in biotin-avidin based assays are well known.  
         [0040]    It is also contemplated within the scope of this invention that the nucleic acid probe assays of this invention will employ a cocktail of nucleic acid probes capable of detecting the gene or genes. Thus, in one example to detect the presence of KVLQT1 in a cell sample, more than one probe complementary to KVLQT1 is employed and in particular the number of different probes is alternatively 2, 3, or 5 different nucleic acid probe sequences. In another example, to detect the presence of mutations in the KVLQT1 gene sequence in a patient, more than one probe complementary to KVLQT1 is employed where the cocktail includes probes capable of binding to the allele-specific mutations identified in populations of patients with alterations in KVLQT1. In this embodiment, any number of probes can be used.  
         [0041]    Large amounts of the polynucleotides of the present invention may be produced by replication in a suitable host cell. Natural or synthetic polynucleotide fragments coding for a desired fragment will be incorporated into recombinant polynucleotide constructs, usually DNA constructs, capable of introduction into and replication in a prokaryotic or eukaryotic cell. Usually the polynucleotide constructs will be suitable for replication in a unicellular host, such as yeast or bacteria, but may also be intended for introduction to (with and without integration within the genome) cultured mammalian or plant or other eukaryotic cell lines. The purification of nucleic acids produced by the methods of the present invention are described, e.g., in Sambrook et al., 1989 or Ausubel et al., 1992.  
         [0042]    The polynucleotides of the present invention may also be produced by chemical synthesis, e.g., by the phosphoramidite method described by Beaucage and Caruthers (1981) or the triester method according to Matteucci and Caruthers (1981) and may be performed on commercial, automated oligonucleotide synthesizers. A double-stranded fragment may be obtained from the single-stranded product of chemical synthesis either by synthesizing the complementary strand and annealing the strand together under appropriate conditions or by adding the complementary strand using DNA polymerase with an appropriate primer sequence.  
         [0043]    Polynucleotide constructs prepared for introduction into a prokaryotic or eukaryotic host may comprise a replication system recognized by the host, including the intended polynucleotide fragment encoding the desired polypeptide, and will preferably also include transcription and translational initiation regulatory sequences operably linked to the polypeptide encoding segment. Expression vectors may include, for example, an origin of replication or autonomously replicating sequence (ARS) and expression control sequences, a promoter, an enhancer and necessary processing information sites, such as ribosome-binding sites, RNA splice sites, polyadenylation sites, transcriptional terminator sequences, and mRNA stabilizing sequences. Such vectors may be prepared by means of standard recombinant techniques well known in the art and discussed, for example, in Sambrook et al. (1989) or Ausubel et al. (1992).  
         [0044]    An appropriate promoter and other necessary vector sequences will be selected so as to be functional in the host, and may include, when appropriate, those naturally associated with the KVLQT1 or other gene. Examples of workable combinations of cell lines and expression vectors are described in Sambrook et al. (1989) or Ausubel et al. (1992); see also, e.g., Metzger et al. (1988). Many useful vectors are known in the art and may be obtained from such vendors as Stratagene, New England Biolabs, Promega Biotech, and others. Promoters such as the trp, lac and phage promoters, tRNA promoters and glycolytic enzyme promoters may be used in prokaryotic hosts. Useful yeast promoters include promoter regions for metallothionein, 3-phosphoglycerate kinase or other glycolytic enzymes such as enolase or glyceraldehyde-3-phosphate dehydrogenase, enzymes responsible for maltose and galactose utilization, and others. Vectors and promoters suitable for use in yeast expression are further described in Hitzeman et al., EP 73,675A. Appropriate non-native mammalian promoters might include the early and late promoters from SV40 (Fiers et al., 1978) or promoters derived from murine Molony leukemia virus, mouse tumor virus, avian sarcoma viruses, adenovirus II, bovine papilloma virus or polyoma. Insect promoters may be derived from baculovirus. In addition, the construct may be joined to an amplifiable gene (e.g., DHFR) so that multiple copies of the gene may be made. For appropriate enhancer and other expression control sequences, see also  Enhancers and Eukaryotic Gene Expression , Cold Spring Harbor Press, Cold Spring Harbor, New York (1983). See also, e.g., U.S. Pat. Nos. 5,691,198; 5,735,500; 5,747,469 and 5,436,146.  
         [0045]    While such expression vectors may replicate autonomously, they may also replicate by being inserted into the genome of the host cell, by methods well known in the art.  
         [0046]    Expression and cloning vectors will likely contain a selectable marker, a gene encoding a protein necessary for survival or growth of a host cell transformed with the vector. The presence of this gene ensures growth of only those host cells which express the inserts. Typical selection genes encode proteins that a) confer resistance to antibiotics or other toxic substances, e.g. ampicillin, neomycin, methotrexate, etc., b) complement auxotrophic deficiencies, or c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli. The choice of the proper selectable marker will depend on the host cell, and appropriate markers for different hosts are well known in the art.  
         [0047]    The vectors containing the nucleic acids of interest can be transcribed in vitro, and the resulting RNA introduced into the host cell by well-known methods, e.g., by injection (see, Kubo et al. (1988)), or the vectors car be introduce 1 directly into host cells by methods well known in the art, which vary depending on the type of cellular host, including electroporation; transfection employing calcium chloride, rubidium chloride calcium phosphate, DEAE-dextran, or other substances; microprojectile bombardment; lipofection; infection (where the vector is an infectious agent, such as a retroviral genome); and other methods. See generally, Sambrook et al. (1989) and Ausubel et al. (1992). The introduction of the polynucleotides into the host cell by any method known in the art, including, inter alia, those described above, will be referred to herein as “transformation.” The cells into which have been introduced nucleic acids described above are meant to also include the progeny of such cells.  
         [0048]    Large quantities of the nucleic acids and polypeptides of the present invention may be prepared by expressing the KVLQT1 nucleic acid or portions thereof in vectors or other expression vehicles in compatible prokaryotic or eukaryotic host cells. The most commonly used prokaryotic hosts are strains of  Escherichia coli , although other prokaryotes, such as Bacillus subtilis or Pseudomonas may also be used.  
         [0049]    Mammalian or other eukaryotic host cells, such as those of yeast, filamentous fungi, plant, insect, or amphibian or avian species, may also be useful for production of the proteins of the present invention. Propagation of mammalian cells in culture is per se well known. See, Jakoby and Pastan (eds.) (1979). Examples of commonly used mammalian host cell lines are VERO and HeLa cells, Chinese hamster ovary (CHO) cells, and WI38, BHK, and COS cell lines, although it will be appreciated by the skilled practitioner that other cell lines may be appropriate, e.g., to provide higher expression, desirable glycosylation patterns, or other features. An example of a commonly used insect cell line is SF9.  
         [0050]    Clones are selected by using markers depending on the mode of the vector construction. The marker may be on the same or a different DNA molecule, preferably the same DNA molecule. In prokaryotic hosts, the transformant may be selected, e.g., by resistance to ampicillin, tetracycline or other antibiotics. Production of a particular product based on temperature sensitivity may also serve as an appropriate marker.  
         [0051]    Prokaryotic or eukaryotic cells transformed with the polynucleotides of the present invention will be useful not only for the production of the nucleic acids and polypeptides of the present invention, but also, for example, in studying the characteristics of KVLQT1 or other polypeptides.  
         [0052]    The probes and primers based on the KVLQT1 or other gene sequences disclosed herein are used to identify homologous KVLQT1 or other gene sequences and proteins in other species. These gene sequences and proteins are used in the diagnostic/prognostic, therapeutic and drug screening methods described herein for the species from which they have been isolated.  
         [0053]    The studies described in the Examples below resulted in the determination of many novel mutations. Previous studies had defined 126 distinct disease causing mutations in the LQTS genes KVLQT1, HERG, SCN5A, KCNE1 and KCNE2 (Wang Q. et al., 1996a; Curran et al., 1995; Wang et al., 1995a; Splawski et al., 1997a; Abbott et al., 1999; Chouabe et al., 1997; Wollnik et al., 1997; Neyroud et al., 1997; Splawski et al., 1997b; Tyson et al., 1997; Schulze-Bahr et al., 1997; Priori et al., 1999; Splawski et al., 1998; Wang et al., 1995b; Russell et al., 1996; Neyroud et al., 1998; Neyroud et al., 1999; Donger et al., 1997; Tanaka et al., 1997; Jongbloed et al., 1999; Priori et al., 1998; Itoh et al., 1998a; Itoh et al., 1998b; Mohammad-Panah et al., 1999; Saarinen et al., 1998; Ackerman et al., 1998; Berthet et al., 1999; Kanters, 1998; van den Berg et al., 1997; Dausse et al., 1996; Benson et al., 1996; Akimoto et al., 1998; Satler et al., 1996; Satler et al., 1998; Makita et al., 1998, An et al., 1998; Schulze-Bahr et al., 1995; Duggal et al., 1998; Chen Q. et al., 1999; Li et al., 1998; Wei et al., 1999; Larsen et al., 1999a; Bianchi et al., 1999; Ackerman et al., 1999a; Ackerman et al., 1999b; Murray et al., 1999; Larsen et al., 1999b; Yoshida et al., 1999; Wattanasirichaigoon et al., 1999; Bezzina et al., 1999; Hoorntje et al., 1999). The sequence of each wild-type gene has been published. The KVLQT1 can be found in Splawski et al. (1998) and the coding region of the cDNA is shown herein as SEQ ID NO:1 and the encoded KVLQT1 is shown as SEQ ID NO:2. SCN5A was reported by Gellens et al. (1992) and its sequence is provided by GenBank Accession No. NM — 000335. The coding sequence of SCN5A is shown herein as SEQ ID NO:3 and the encoded SCN5A is shown as SEQ ID NO:4. Most of the mutations were found in KVLQT1 (Yoshida et al., 1999) and HERG (Itoh et al., 1998b), and fewer in SCNSA (Wang Q. et al., 1996a), KCNEI (Jiang et al., 1994) and KCNE2 (Ward, 1964). These mutations were identified in regions with known intron/exon structure, primarily the transmembrane and pore domains. In this study, we screened 262 individuals with LQTS for mutations in all known arrhythmia genes. We identified 134 mutations, 80 of which were novel. Together with 43 mutations reported in our previous studies, we have now identified 177 mutations in these 262 LQTS individuals (68%). The failure to identify mutations in 32% of the individuals may result from phenotypic errors, incomplete sensitivity of SSCP or presence of mutations in regulatory sequences. However, it is also clear that additional LQTS genes await discovery (Jiang et al., 1994; Schott et al., 1995).  
         [0054]    Missense mutations were most common (72%), followed by frameshift mutations (10%), in-frame deletions, nonsense and splice site mutations (5-7% each). Most mutations resided in intracellular (52%) and transmembrane (30%) domains; 12% were found in pore and 6% in extracellular segments. One hundred one of the 129 distinct LQTS mutations (78%) were identified in single families or individuals. Most of the 177 mutations were found in KVLQTI (75 or 42%) and HERG (80 or 45%). These two genes accounted for 87% of the identified mutations, while mutations in SCN5A (14 or 8%), KCNE1 (5 or 3%) and KCNE2 (3 or 2%) accounted for the other 13%.  
         [0055]    Multiple mutations were found in regions encoding S5, S5/P, P and S6 of KVLQT1 and HERG. The P region of potassium channels forms the outer pore and contains the selectivity filter (Doyle et al., 1998). Transmembrane segment 6, corresponding to the inner helix of KcsA, forms the inner 2/3 of the pore. This structure is supported by the S5 transmembrane segment, corresponding to the outer helix of KcsA, and is conserved from prokaryotes to eukaryotes ((MacKinnon et al., 1998). Mutations in these regions will likely disrupt potassium transport. Many mutations were identified in the C-termini of KVLQT1 and HERG. Changes in the C-terminus of HERG could lead to anomalies in tetramerization as it has been proposed that the C-terminus of eag, which is related to HERG, is involved in this process (Ludwig et al, 1994).  
         [0056]    Multiple mutations were also identified in regions that were different for KVLQT1 and HERG. In KVLQT1, multiple mutations were found in the sequences coding for the S2/S3 and S4/S5 linkers. Coexpression of S2/S3 mutants with wild-type KVLQT1 in  Xenopus oocytes  led to simple loss of function or dominant-negative effect without significantly changing the biophysical properties of I Ks  channels (Chouabe et al., 1997; Shalaby et al., 1997; Wang et al., 1999). On the other hand, S4/S5 mutations altered the gating properties of the channels and modified KVLQT1 interactions with minK subunits (Wang et al., 1999; Franqueza et al., 1999). In HERG, more than 20 mutations were identified in the N-terminus. HERG channels lacking this region deactivate faster and mutations in the region had a similar effect (Chen J. et al., 1999).  
         [0057]    Mutations in KCNE1 and KCNE2, encoding minK and MiRP1, the respective I Ks  and I Kr  β-subunits, altered the biophysical properties of the channels (Splawski et al., 1997a; Abbott et al., 1999; Sesti and Goldstein, 1998). A Mir P1 mutant, involved in clarithromyocin-induced arrhythmia, increased channel blockade by the antibiotic (Abbott et al., 1999). Mutations in SCN5A, the sodium channel α-subunit responsible for cardiac I Na , destabilized the inactivation gate causing delayed channel inactivation and dispersed reopenings (Bennett et al., 1995; Dumaine et al., 1996; Wei et al., 1999; Wang D W et al., 1996). One SCN5A mutant affected the interactions with the sodium channel β-subunit (An et al., 1998).  
         [0058]    It is interesting to note that probands with KCNE1 and KCNE2 mutations were older and had shorter QTc than probands with the other genotypes. The significance of these differences is unknown, however, as the number of probands with KCNE1 and KCNE2 genotypes was small.  
         [0059]    This catalogue of mutations will facilitate genotype-phenotype analyses. It also has clinical implications for presymptomatic diagnosis and, in some cases, for therapy. Patients with mutations in KVLQT1, HERG, KCNE1 and KCNE2, for example, may benefit from potassium therapy (Compton et al., 1996). Sodium channel blockers, on the other hand, might be helpful in patients with SCN5A mutations (Schwartz et al. (1995). The identification of mutations is of importance for ion channel studies as well. The expression of mutant channels in heterologous systems can reveal how structural changes influence the behavior of the channel or how mutations affect processing (Zhou et al., 1998; Furutani et al., 1999). These studies improve our understanding of channel function and provide insights into mechanisms of disease. Finally, mutation identification will contribute to the development of genetic screening for arrhythmia susceptibility.  
         [0060]    The present invention is described by reference to the following Examples, which are offered by way of illustration and are not intended to limit the invention in any manner. Standard techniques well known in the art or the techniques specifically described in the Examples were utilized.  
       EXAMPLE 1  
     Ascertainment and Phenotyping  
       [0061]    Individuals were ascertained in clinics from North America and Europe. Individuals were evaluated for LQTS based on QTc (the QT interval corrected for heart rate) and for the presence of symptoms. In this study, we focused on the probands. Individuals show prolongation of the QT interval (QTc≧460 ms) and/or documented torsade depointes, ventricular fibrillation, cardiac arrest or aborted sudden death. Informed consent was obtained in accordance with local institutional review board guidelines. Phenotypic data were interpreted without knowledge of genotype. Sequence changes altering coding regions or predicted to affect splicing that were not detected in at least 400 control chromosomes were defined as mutations. No changes except known polymorphisms were detected ina ny of the genes in the control population. This does not exclude the possibility that some mutations are rare variants not associated with disease.  
       EXAMPLE 2  
     Mutational Analyses  
       [0062]    To determine the spectrum of LQTS mutations, we used SSCP (Single Stand Conformation Polymorphism) and DNA sequence analyses to screen 262 unrelated individuals with LQTS. Seventeen primer pairs were used to screen KVLQT1 (Splawski et al., 1998), twenty-one primer pairs were used for HERG (Splawski et al., 1998) and three primer pairs were used for KCNE1 (Splawski et al., 1997a) and KCNE2 (Abbott et al., 1999). Thirty-three primer pairs (Wang Q. et al., 1996b) were used in SSCP analysis to screen all SCN5A exons in 50 individuals with suspected abnormalities in I Na . Exons 23-28, in which mutations were previously identified, were screened in all 262 individuals.  
         [0063]    Gender, age, QTc and presence of symptoms are summarized in Table 1. The average age at ascertainment was 29 with a corrected QT interval of 492 ms. Seventy-five percent had a history of symptoms and females predominated with an ˜2:1 ratio. Although the numbers were small, corrected QT intervals for individuals harboring KCNE1 and KCNE2 mutations were shorter at 457 ms.  
                                     TABLE 1                           Age, QTc, Gender and Presence of Symptoms                Age*, y       QTc, ms           Genotype   (mean ± SD)   Gender (F/M)   (mean ± SD)   Symptoms †                 KVLQT1   32 ± 19   52/23   493 ± 45   78%       HERG   31 ± 19   51/29   498 ± 48   71%       SCN5A   32 ± 24   8/6   511 ± 42   55%       KCNE1   43 ± 16   3/2   457 ± 25   40%       KCNE2   54 ± 20   3/0   457 ± 05   67%       unknown   25 ± 16   56/29   484 ± 46   81%       all   29 ± 19   173/89   492 ± 47   75%                                  
 
         [0064]    The SSCP analyses revealed many mutations. KVLQT1 mutations associated with LQTS were identified in 52 individuals (FIG. 1 and Table 2). Twenty of the mutations were novel. HERG mutations were identified in 68 LQTS individuals (FIG. 2 and Table 3). Fifty-two of these mutations were novel. SCN5A mutations were identified in eight cases (FIG. 3 and Table 4). Five of the mutations were novel. Three novel KCNE1 mutations were identified (FIG. 4 and Table 5) and three mutations were identified in KCNE2 FIG. 5 and Table 6) (Abbott et al., 1999). None of the KVLQT1, HERG, SCN5A, KCNE1 and KCNE2 mutations was observed in 400 control chromosomes.  
                                                                   TABLE 2                           Summary of All KVLQT1 Mutations*                            Number           Nucleotide   Coding           of       Change†   Effect   Position   Exon   families‡   Study                    del211-219   del71-73   N-terminus   1   1   Ackerman                           et al., 1999a       A332G†   Y111C   N-terminus   1   1   This       del451-452   A150fs/132   S2   2   1 JLN   Chen Q.                           et al., 1999       T470G   F157C   S2   1   1   Larsen et al.,                           1999a       G477 + 1A   M159sp   S2   2   1 JLN,   This; Donger                       1 UK   et al., 1997       G477 + 5A   M159sp   S2   1   1   Ackerman                           et al., 1999b       G478A†   E160K   S2   3   1   This       del500-502   F167W/del   S2   3   1   Wang Q.           G168               et al., 1996a       G502A   G168R   S2   3   7   This;                           Splawski                           et al., 1998;                           Donger                           et al., 1997       C520T   R174C   S2/S3   3   1   Donger                           et al, 1997       G521A†   R174H   S2/S3   3   1   This       G532A   A178T   S2/S3   3   1   Tanaka                           et al., 1997       G532C   A178P   S2/S3   3   1   Wang Q.                           et al., 1996a       G535A†   G179S   S2/S3   3   1   This       A551C   Y184S   S2/S3   3   2   This;                           Jongbloed                           et al., 1999       G565A   G189R   S2/S3   3   3   Wang Q.                           et al., 1996a                           Jongbloed                           et al., 1999       insG567-   G189fs/94   S2/S3   3   1   Splawski       568               (RW +   et al., 1997b                       JLN)       G569A   R190Q   S2/S3   3   2   Splawski                           et al., 1998;                           Donger                           et al., 1997       del572-576   L191fs/90   S2/S3   3   1 JLN,   Tyson et al.,                       1 RW   1997;                       2   Ackerman                       (JLN +   et al., 1999b                       RW)       G580C†   A194P   S2/S3   3   1   This       C674T   S225L   S4   4   2   This; Priori                           et al., 1999       G724A   D242N   S4/S5   5   1   Itoh et al.,                           1998b       C727T†   R243C   S4/S5   5   2   This       G728A   R243H   S4/S5   5   1 JLN   Saarinen                           et al., 1998       T742C†   W248R   S4/S5   5   1   This       T749A   L250H   S4/S5   5   1   Itoh et al.,                           1998a       G760A   V254M   S4/S5   5   4   This;                           Wang Q.                           et al.,                           1996A;                           Donger                           et al., 1997       G781A   E261K   S4/S5   6   1   Donger                           et al., 1997       T797C†   L266P   S5   6   1   This       G805A   G269S   S5   6   1   Ackerman                           et al., 1999b       G806A   G269D   S5   6   3   This;                           Donger                           et al., 1997       C817T   L273F   S5   6   2   This;                           Wang Q.                           et al., 1996a       A842G   Y281C   S5   6   1   Priori et al.,                           1999       G898A   A300T   S5/Pore   6   1   Priori et al.,                           1998       G914C   W305S   Pore   6   1 JLN   Chouabe                           et al., 1997       G916A   G306R   Pore   6   1   Wang Q.                           et al, 1996a       del921 −   V307sp   Pore   6   1   Li et al.,                           1998       (921 + 2)       G921 + 1T†   V307sp   Pore   6   1   This       A922 − 2C†   V307sp   Pore   7   1   This       G922 − 1C   V307sp   Pore   7   1   Murray                           et al., 1999       C926G   T309R   Pore   7   1   Donger                           et al., 1997       G928A†   V310I   Pore   7   1   This       C932T   T311I   Pore   7   1   Saarinen                           et al., 1998       C935T   T312I   Pore   7   2   This;                           Wang Q.                           et al., 1996a       C939G   I313M   Pore   7   1   Tanaka                           et al., 1997       G940A   G314S   Pore   7   7   Splawski                           et al., 1998;                           Russell                           et al., 1996;                           Donger                           et al., 1997;                           Jongbloed                           et al., 1999;                           Itoh et al.,                           1998b       A944C   Y315S   Pore   7   3   Donger                           et al., 1997;                           Jongbloed                           et al., 1999       A944G   Y315C   Pore   7   2   Priori et al.,                           1999;                           Splawski                           et al., 1998       G949A   D317N   Pore   7   2   Wollnik                           et al., 1997;                           Saarinen                           et al., 1998       G954C   K318N   Pore   7   1   Splawski                           et al., 1998       C958G   P320A   Pore   7   1   Donger                           et al., 1997       G973A   G325R   S6   7   4   This; Donger                           et al., 1997;                           Tanaka                           et al., 1997       del1017-   delF340   S6   7   2   This;       1019                    Ackerman                           et al., 1998       C1022A   A341E   S6   7   5   This;                           Wang Q.                           et al., 1996a;                           Berthet                           et al., 1999       C1022T   A341V   S6   7   7   This;                           Wang Q.                           et al., 1996a;                           Russell                           et al., 1996;                           Donger                           et al., 1997;                           Li et al.,                           1998       C1024T   L342F   S6   7   1   Donger                           et al., 1997       C1031T   A344V   S6   7   1   Donger                           et al., 1997       G1032A   A344sp   S6   7   9   This;                           Kanters,                           1998; Li et                           al., 1998;                           Ackerman                           et al., 1999b;                           Murray                           et al., 1999       G1032C   A344sp   S6   7   1   Murray                           et al., 1999       G1033C   G345R   S6   8   1   van den Berg                           et al., 1997       G1034A   G345E   S6   8   1   Wang Q.                           et al., 1996a       C1046G†   S349W   S6   8   1   This       T1058C   L353P   S6   8   1   Splawski                           et al., 1998       C1066T†   Q356X   C-terminus   8   1   This       C1096T   R366W   C-terminus   8   1   Splawski                           et al., 1998       G1097A†   R366Q   C-terminus   8   1   This       G1097C   R366P   C-terminus   8   1   Tanaka                           et al., 1997       G1111A   A371T   C-terminus   8   1   Donger                           et al., 1997       T1117C   S373P   C-terminus   8   1   Jongbloed                           et al., 1999       C1172T†   T391I   C-terminus   9   1   This       T1174C   W392R   C-terminus   9   1   Jongbloed                           et al., 1999       C1343G†   P448R   C-terminus   10   2   This       C1522T   R518X   C-terminus   12   1 JLN,   This; Larsen                       3 RW   et al., 1999       G1573A   A525T   C-terminus   12   1   Larsen et al.,                           1999b       C1588T†   Q530X   C-terminus   12   1 JLN,   This                       1 RW       C1615T   R539W   C-terminus   13   1   Chouabe                           et al., 1997       del6/ins7   E543fs/107   C-terminus   13   1 JLN   Neyroud                           et al., 1997       C1663T   R555C   C-terminus   13   3   Donger                           et al., 1997       C1697T†   S566F   C-terminus   14   3   This       C1747T†   R583C   C-terminus   15   1   This       C1760T   T587M   C-terminus   15   1 JLN,   Donger                       1 RW   et al., 1997,                           Itoh et al.,                           1998b       G1772A   R591H   C-terminus   15   1   Donger                           et al., 1997       G1781A†   R594Q   C-terminus   15   3   This       del1892-   P630fs/13   C-terminus   16   1 JLN   Donger       1911                   et al., 1997       insC1893-   P631fs/19   C-terminus   16   1   Donger       1894                   et al, 1997                                          
 
         [0065]    [0065]                                                                   TABLE 3                           Summary of All HERG Mutations*                            Number           Nucleotide   Coding           of RW       Change   Effect   Position   Exon   Families   Study                    C87A†   F29L   N-terminus   2   1   This       A98C†   N33T   N-terminus   2   2   This       C132A†   C44X   N-terminus   2   1   This       G140T†   G47V   N-terminus   2   1   This       G157C†   G53R   N-terminus   2   1   This       G167A†   R56Q   N-terminus   2   1   This       T196G†   C66G   N-terminus   2   1   This       A209G†   H70R   N-terminus   2   2   This       C215A†   P72Q   N-terminus   2   2   This       del221-251†   R73sf/31   N-terminus   2   1   This       G232C†   A78P   N-terminus   2   1   This       dupl234-   A83fs/37   N-terminus   2   1   This       250†       C241T†   Q81X   N-terminus   2   1   This       T257G†   L86R   N-terminus   2   1   This       insC422-   P141sf/2   N-terminus   3   1   This       423†       insC453-   P151fs/   N-terminus   3   1   This       454†   179       dupl558-600   L200sf/   N-terminus   4   1   Hoorntje           144               et al., 1999       insC724-   P241fs/89   N-terminus   4   1   This       725†       del885†   V295fs/63   N-terminus   4   1   This       C934T†   R312C   N-terminus   5   1   This       C1039T†   P347S   N-terminus   5   1   This       G1128A†   Q376sp   N-terminus   5   1   This       A1129-2G†   Q376sp   N-terminus   6   1   This       del1261   Y420fs/12   S1   6   1   Curran et al.,                           1995       C1283A   S428X   S1/S2   6   1   Priori et al.,                           1999       C1307T   T436M   S1/S2   6   1   Priori et al.,                           1999       A1408G   N470D   S2   6   1   Curran et al,                           1995       C1421T   T474I   S2/S3   6   1   Tanaka et                           al., 1997       C1479G   Y493X   S2/S3   6   1   Itoh et al.,                           1998a       del1498-   del500-   S3   6   1   Curran et al.,       1524   508               1995       G1592A†   R531Q   S4   7   1   This       C1600T   R534C   S4   7   1   Itoh et al.,                           1998a       T1655C†   L552S   S5   7   1   This       delT1671   T556fs/7   S5   7   1   Schulze-                           Bahr et al.,                           1995       G1672C   A558P   S5   7   1   Jongbloed                           et al., 1999       G1681A   A561T   S5   7   4   This;                           Dausse                           et al., 1996       C1682T   A561V   S5   7   4   This; Curran                           et al., 1995;                           Priori et al.,                           1999       G1714C   G572R   S5/Pore   7   1   Larsen et al.,                           1999a       G1714T   G572C   S5/Pore   7   1   Splawski                           et al., 1998       C1744T   R582C   S5/Pore   7   1   Jongbloed                           et al., 1999       G1750A†   G584S   S5/Pore   7   1   This       G1755T†   W585C   S5/Pore   7   1   This       A1762G   N588D   S5/Pore   7   1   Splawski                           et al., 1998       T1778C†   I593T   S5/Pore   7   1   This       T1778G   I593R   S5/Pore   7   1   Benson                           et al., 1996       G1801A   G601S   S5/Pore   7   1   Akimoto                           et al., 1998       G1810A   G604S   S5/Pore   7   2   This;                           Jongbloed                           et al., 1999       G1825A†   D609N   S5/Pore   7   1   This       T1831C   Y611H   S5/Pore   7   1   Tanaka                           et al., 1997       T1833 (A or   Y611X   S5/Pore   7   1   Schulze-       G)                   Bahr et al.,                           1995       G1834T   V612L   Pore   7   1   Satler et al.,                           1998       C1838T   T613M   Pore   7   4   This;                           Jongbloed                           et al., 1999       C1841T   A614V   Pore   7   6   Priori et al.,                           1999;                           Splawski et                           al., 1998;                           Tanaka et                           al., 1997;                           Satler et al.,                           1998       C1843G†   L615V   Pore   7   1   This       G1876A†   G626S   Pore   7   1   This       C1881G†   F627L   Pore   7   1   This       G1882A   G628S   Pore   7   2   This; Curran                           et al., 1995       A1885G   N629D   Pore   7   1   Satler et al.,                           1998       A1886G   N629S   Pore   7   1   Satler et al.,                           1998       C1887A   N629K   Pore   7   1   Yoshida et                           al., 1999       G1888C   V630L   Pore   7   1   Tanaka et                           al., 1997       T1889C   V630A   Pore   7   1   Splawski et                           al., 1998       C1894T†   P632S   Pore   7   1   This       A1898G   N633S   Pore   7   1   Satler et al.,                           1998       A1912G†   K638E   S6   7   1   This       del1913-   delK638   S6   7   1   This       1915†       C1920A   F640L   S6   7   1   Jongbloed                           et al., 1999       A1933T†   M645L   S6   7   1   This       del1951-   L650fs/2   S6   8   1   Itoh et al.,       1952                   1998a       G2044T†   E682X   S6/cNBD   8   1   This       C2173T   Q725X   S6/cNBD   9   1   Itoh et al.,                           1998a       insT2218-   H739fs/63   S6/cNBD   9   1   This       2219†       C2254T†   R752W   S6/cNBD   9   1   This       dupl2356-   V796fs/22   cNBD   9   1   Itoh et al.,       2386                   1998a       del2395†   I798fs/10   cNBD   9   1   This       G2398 + 1C   L799sp   cNBD   9   2   This;                           Curran et al.,                           1995       T2414C†   F805S   cNBD   10   1   This       T2414G†   F805C   cNBD   10   1   This       C2453T   S818L   cNBD   10   1   Berthet                           et al., 1999       G2464A   V822M   cNBD   10   2   Berthet                           et al., 1999;                           Satler et al.,                           1996       C2467T†   R823W   cNBD   10   2   This       A2582T†   N861I   C-terminus   10   1   This       G2592 + 1A   D864sp   C-terminus   10   2   This; Berthet                           et al., 1999       del2660†   K886fs/85   C-terminus   11   1   This       C2750T†   P917L   C-terminus   12   1   This       del2762†   R920fs/51   C-terminus   12   1   This       C2764T†   R922W   C-terminus   12   1   This       insG2775-   G925fs/13   C-terminus   12   1   This       2776†       del2906†   P968fs/4   C-terminus   12   1   This       del2959-   P986fs/   C-terminus   12   1   This       2960†   130       C3040T†   R1014X   C-terminus   13   2   This       del3094†   G1031fs/   C-terminus   13   1   This           24       insG3107-   G1036fs/   C-terminus   13   1   Berthet       3108   82               et al., 1999       insC3303-   P1101fs   C-terminus   14   1   This       3304†                            
         [0066]    [0066]                                         TABLE 4                           Summary of All SCN5A Mutations                            Number of           Nucleotide Change   Coding Effect   Position   Exon   RW Families   Study               G3340A†   D1114N   DII/DIII   18   1   This       C3911T   T1304M   DIII/S4   22   1   Wattanasirichaigoon et al.,                           1999       A3974G   N1325S   DIII/S4/S5   23   1   Wang et al., 1995b       C4501G†   L1501V   DIII/DIV   26   1   This       del4511-   del1505-   DIII/DIV   26   4   Wang et al., 1995a; Wang et       4519   1507               al., 1995b       del4850-   delF1617   DIV/S3/S4   28   1   This       4852†       G4868A   R1623Q   DIV/S4   28   2   This; Makita et al., 1998       G4868T†   R1623L   DIV/S4   28   1   This       G4931A   R1644H   DIV/S4   28   2   This; Wang et al., 1995b       C4934T   T1645M   DIV/S4   28   1   Wattanasirichaigoon et al.,                           1999       G5350A†   E1784K   C-terminus   28   2   This; Wei et al., 1999       G536OA†   S1787N   C-terminus   28   1   This       A5369G   D1790G   C-terminus   28   1   An et al., 1998       insTGA   insD1795-   C-terminus   28   1   Bezzina et al., 1999       5385-5386   1796                            
         [0067]    [0067]                                         TABLE 5                           Summary of All KCNE1 Mutations*                            Number           Nucleotide   Coding           of       Change   Effect   Position   Exon   Families   Study               C20T   T7I   N-terminus   3   1 JLN   Schulze-                           Bahr et al.,                           1997       G95A†   R32H   N-terminus   3   1   This       G139T   V47F   S1   3   1 JLN   Bianchi et                           al., 1999       TG151-   L51H   S1   3   1 JLN   Bianchi et       152AT                   al., 1999       A172C/TG   TL58-   S1   3   1 JLN   Tyson et al.,       176-177CT   59PP               1997       C221T   S74L   C-terminus   3   1   Splawski et                           al., 1997a       G226A   D76N   C-terminus   3   1 JLN,   Splawski et                       1 RW,   al., 1997a;                       1 (JLN +   Tyson et al.,                       RW)   1997;                           Duggal et                           al., 1998       T259C   W87R   C-terminus   3   1   Bianchi et                           al., 1999       C292T†   R98W   C-terminus   3   1   This       C379A†   P127T   C-terminus   3   1   This                            
         [0068]    [0068]                                         TABLE 6                           Summary of All KCNE2 Mutations                            Number           Nucleotide   Coding           of       Change   Effect   Position   Exon   Families   Study               C25G   Q9E   N-   1   1   Abbott et al., 1999               terminus       T161T   M54T   S1   1   1   Abbott et al., 1999       T170C   I57T   S1   1   1   Abbott et al., 1999                    
         [0069]    [0069]                                             TABLE 7                           Mutations by Type            Type   KVLQT1   HERG   SCN5A   KCNE1   KCNE2   Total               Missense   59    52    9   5   3   128        Nonsense   6   5   0   0   0   11       AA deletion*   2   2   5   0   0    9       Frameshift   1   16    0   0   0   17       Splice   7   5   0   0   0   12       Total   75    80    14    5   3   177                             
         [0070]    [0070]                                             TABLE 8                           Mutations by Position            Gene                               Protein   KVLQT1   HERG   SCN5A   KCNE1   KCNE2       Position   KVLQT1   HERG   SCN5A   minK   MiRP1   Total               Extracellular    0    7   1   1   1   10       Trans-   33   13   5   0   2   53       membrane       Pore    9   12   0   N/A   N/A   21       Intracellular   33   48   8   4   0   93       Total   75   80   14    5   3   177                     
         [0071]    While the invention has been disclosed in this Patent application by reference to the details of preferred embodiments of the invention, it is to be understood that the disclosure is intended in an illustrative rather than in a limiting sense, as it is contemplated that modifications will readily occur to those skilled in the art, within the spirit of the invention and the scope of the appended claims.  
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         1 
         
           
             4  
           
           
             1  
             2028  
             DNA  
             Homo sapiens  
             
               CDS  
               (1)..(2028)  
             
           
            1 

atg gcc gcg gcc tcc tcc ccg ccc agg gcc gag agg aag cgc tgg ggt       48 
Met Ala Ala Ala Ser Ser Pro Pro Arg Ala Glu Arg Lys Arg Trp Gly 
  1               5                  10                  15 

tgg ggc cgc ctg cca ggc gcc cgg cgg ggc agc gcg ggc ctg gcc aag       96 
Trp Gly Arg Leu Pro Gly Ala Arg Arg Gly Ser Ala Gly Leu Ala Lys 
             20                  25                  30 

aag tgc ccc ttc tcg ctg gag ctg gcg gag ggc ggc ccg gcg ggc ggc      144 
Lys Cys Pro Phe Ser Leu Glu Leu Ala Glu Gly Gly Pro Ala Gly Gly 
         35                  40                  45 

gcg ctc tac gcg ccc atc gcg ccc ggc gcc cca ggt ccc gcg ccc cct      192 
Ala Leu Tyr Ala Pro Ile Ala Pro Gly Ala Pro Gly Pro Ala Pro Pro 
     50                  55                  60 

gcg tcc ccg gcc gcg ccc gcc gcg ccc cca gtt gcc tcc gac ctt ggc      240 
Ala Ser Pro Ala Ala Pro Ala Ala Pro Pro Val Ala Ser Asp Leu Gly 
 65                  70                  75                  80 

ccg cgg ccg ccg gtg agc cta gac ccg cgc gtc tcc atc tac agc acg      288 
Pro Arg Pro Pro Val Ser Leu Asp Pro Arg Val Ser Ile Tyr Ser Thr 
                 85                  90                  95 

cgc cgc ccg gtg ttg gcg cgc acc cac gtc cag ggc cgc gtc tac aac      336 
Arg Arg Pro Val Leu Ala Arg Thr His Val Gln Gly Arg Val Tyr Asn 
            100                 105                 110 

ttc ctc gag cgt ccc acc ggc tgg aaa tgc ttc gtt tac cac ttc gcc      384 
Phe Leu Glu Arg Pro Thr Gly Trp Lys Cys Phe Val Tyr His Phe Ala 
        115                 120                 125 

gtc ttc ctc atc gtc ctg gtc tgc ctc atc ttc agc gtg ctg tcc acc      432 
Val Phe Leu Ile Val Leu Val Cys Leu Ile Phe Ser Val Leu Ser Thr 
    130                 135                 140 

atc gag cag tat gcc gcc ctg gcc acg ggg act ctc ttc tgg atg gag      480 
Ile Glu Gln Tyr Ala Ala Leu Ala Thr Gly Thr Leu Phe Trp Met Glu 
145                 150                 155                 160 

atc gtg ctg gtg gtg ttc ttc ggg acg gag tac gtg gtc cgc ctc tgg      528 
Ile Val Leu Val Val Phe Phe Gly Thr Glu Tyr Val Val Arg Leu Trp 
                165                 170                 175 

tcc gcc ggc tgc cgc agc aag tac gtg ggc ctc tgg ggg cgg ctg cgc      576 
Ser Ala Gly Cys Arg Ser Lys Tyr Val Gly Leu Trp Gly Arg Leu Arg 
            180                 185                 190 

ttt gcc cgg aag ccc att tcc atc atc gac ctc atc gtg gtc gtg gcc      624 
Phe Ala Arg Lys Pro Ile Ser Ile Ile Asp Leu Ile Val Val Val Ala 
        195                 200                 205 

tcc atg gtg gtc ctc tgc gtg ggc tcc aag ggg cag gtg ttt gcc acg      672 
Ser Met Val Val Leu Cys Val Gly Ser Lys Gly Gln Val Phe Ala Thr 
    210                 215                 220 

tcg gcc atc agg ggc atc cgc ttc ctg cag atc ctg agg atg cta cac      720 
Ser Ala Ile Arg Gly Ile Arg Phe Leu Gln Ile Leu Arg Met Leu His 
225                 230                 235                 240 

gtc gac cgc cag gga ggc acc tgg agg ctc ctg ggc tcc gtg gtc ttc      768 
Val Asp Arg Gln Gly Gly Thr Trp Arg Leu Leu Gly Ser Val Val Phe 
                245                 250                 255 

atc cac cgc cag gag ctg ata acc acc ctg tac atc ggc ttc ctg ggc      816 
Ile His Arg Gln Glu Leu Ile Thr Thr Leu Tyr Ile Gly Phe Leu Gly 
            260                 265                 270 

ctc atc ttc tcc tcg tac ttt gtg tac ctg gct gag aag gac gcg gtg      864 
Leu Ile Phe Ser Ser Tyr Phe Val Tyr Leu Ala Glu Lys Asp Ala Val 
        275                 280                 285 

aac gag tca ggc cgc gtg gag ttc ggc agc tac gca gat gcg ctg tgg      912 
Asn Glu Ser Gly Arg Val Glu Phe Gly Ser Tyr Ala Asp Ala Leu Trp 
    290                 295                 300 

tgg ggg gtg gtc aca gtc acc acc atc ggc tat ggg gac aag gtg ccc      960 
Trp Gly Val Val Thr Val Thr Thr Ile Gly Tyr Gly Asp Lys Val Pro 
305                 310                 315                 320 

cag acg tgg gtc ggg aag acc atc gcc tcc tgc ttc tct gtc ttt gcc     1008 
Gln Thr Trp Val Gly Lys Thr Ile Ala Ser Cys Phe Ser Val Phe Ala 
                325                 330                 335 

atc tcc ttc ttt gcg ctc cca gcg ggg att ctt ggc tcg ggg ttt gcc     1056 
Ile Ser Phe Phe Ala Leu Pro Ala Gly Ile Leu Gly Ser Gly Phe Ala 
            340                 345                 350 

ctg aag gtg cag cag aag cag agg cag aag cac ttc aac cgg cag atc     1104 
Leu Lys Val Gln Gln Lys Gln Arg Gln Lys His Phe Asn Arg Gln Ile 
        355                 360                 365 

ccg gcg gca gcc tca ctc att cag acc gca tgg agg tgc tat gct gcc     1152 
Pro Ala Ala Ala Ser Leu Ile Gln Thr Ala Trp Arg Cys Tyr Ala Ala 
    370                 375                 380 

gag aac ccc gac tcc tcc acc tgg aag atc tac atc cgg aag gcc ccc     1200 
Glu Asn Pro Asp Ser Ser Thr Trp Lys Ile Tyr Ile Arg Lys Ala Pro 
385                 390                 395                 400 

cgg agc cac act ctg ctg tca ccc agc ccc aaa ccc aag aag tct gtg     1248 
Arg Ser His Thr Leu Leu Ser Pro Ser Pro Lys Pro Lys Lys Ser Val 
                405                 410                 415 

gtg gta aag aaa aaa aag ttc aag ctg gac aaa gac aat ggg gtg act     1296 
Val Val Lys Lys Lys Lys Phe Lys Leu Asp Lys Asp Asn Gly Val Thr 
            420                 425                 430 

cct gga gag aag atg ctc aca gtc ccc cat atc acg tgc gac ccc cca     1344 
Pro Gly Glu Lys Met Leu Thr Val Pro His Ile Thr Cys Asp Pro Pro 
        435                 440                 445 

gaa gag cgg cgg ctg gac cac ttc tct gtc gac ggc tat gac agt tct     1392 
Glu Glu Arg Arg Leu Asp His Phe Ser Val Asp Gly Tyr Asp Ser Ser 
    450                 455                 460 

gta agg aag agc cca aca ctg ctg gaa gtg agc atg ccc cat ttc atg     1440 
Val Arg Lys Ser Pro Thr Leu Leu Glu Val Ser Met Pro His Phe Met 
465                 470                 475                 480 

aga acc aac agc ttc gcc gag gac ctg gac ctg gaa ggg gag act ctg     1488 
Arg Thr Asn Ser Phe Ala Glu Asp Leu Asp Leu Glu Gly Glu Thr Leu 
                485                 490                 495 

ctg aca ccc atc acc cac atc tca cag ctg cgg gaa cac cat cgg gcc     1536 
Leu Thr Pro Ile Thr His Ile Ser Gln Leu Arg Glu His His Arg Ala 
            500                 505                 510 

acc att aag gtc att cga cgc atg cag tac ttt gtg gcc aag aag aaa     1584 
Thr Ile Lys Val Ile Arg Arg Met Gln Tyr Phe Val Ala Lys Lys Lys 
        515                 520                 525 

ttc cag caa gcg cgg aag cct tac gat gtg cgg gac gtc att gag cag     1632 
Phe Gln Gln Ala Arg Lys Pro Tyr Asp Val Arg Asp Val Ile Glu Gln 
    530                 535                 540 

tac tcg cag ggc cac ctc aac ctc atg gtg cgc atc aag gag ctg cag     1680 
Tyr Ser Gln Gly His Leu Asn Leu Met Val Arg Ile Lys Glu Leu Gln 
545                 550                 555                 560 

agg agg ctg gac cag tcc att ggg aag ccc tca ctg ttc atc tcc gtc     1728 
Arg Arg Leu Asp Gln Ser Ile Gly Lys Pro Ser Leu Phe Ile Ser Val 
                565                 570                 575 

tca gaa aag agc aag gat cgc ggc agc aac acg atc ggc gcc cgc ctg     1776 
Ser Glu Lys Ser Lys Asp Arg Gly Ser Asn Thr Ile Gly Ala Arg Leu 
            580                 585                 590 

aac cga gta gaa gac aag gtg acg cag ctg gac cag agg ctg gca ctc     1824 
Asn Arg Val Glu Asp Lys Val Thr Gln Leu Asp Gln Arg Leu Ala Leu 
        595                 600                 605 

atc acc gac atg ctt cac cag ctg ctc tcc ttg cac ggt ggc agc acc     1872 
Ile Thr Asp Met Leu His Gln Leu Leu Ser Leu His Gly Gly Ser Thr 
    610                 615                 620 

ccc ggc agc ggc ggc ccc ccc aga gag ggc ggg gcc cac atc acc cag     1920 
Pro Gly Ser Gly Gly Pro Pro Arg Glu Gly Gly Ala His Ile Thr Gln 
625                 630                 635                 640 

ccc tgc ggc agt ggc ggc tcc gtc gac cct gag ctc ttc ctg ccc agc     1968 
Pro Cys Gly Ser Gly Gly Ser Val Asp Pro Glu Leu Phe Leu Pro Ser 
                645                 650                 655 

aac acc ctg ccc acc tac gag cag ctg acc gtg ccc agg agg ggc ccc     2016 
Asn Thr Leu Pro Thr Tyr Glu Gln Leu Thr Val Pro Arg Arg Gly Pro 
            660                 665                 670 

gat gag ggg tcc                                                     2028 
Asp Glu Gly Ser 
        675 

 
           
             2  
             676  
             PRT  
             Homo sapiens  
           
            2 

Met Ala Ala Ala Ser Ser Pro Pro Arg Ala Glu Arg Lys Arg Trp Gly 
  1               5                  10                  15 

Trp Gly Arg Leu Pro Gly Ala Arg Arg Gly Ser Ala Gly Leu Ala Lys 
             20                  25                  30 

Lys Cys Pro Phe Ser Leu Glu Leu Ala Glu Gly Gly Pro Ala Gly Gly 
         35                  40                  45 

Ala Leu Tyr Ala Pro Ile Ala Pro Gly Ala Pro Gly Pro Ala Pro Pro 
     50                  55                  60 

Ala Ser Pro Ala Ala Pro Ala Ala Pro Pro Val Ala Ser Asp Leu Gly 
 65                  70                  75                  80 

Pro Arg Pro Pro Val Ser Leu Asp Pro Arg Val Ser Ile Tyr Ser Thr 
                 85                  90                  95 

Arg Arg Pro Val Leu Ala Arg Thr His Val Gln Gly Arg Val Tyr Asn 
            100                 105                 110 

Phe Leu Glu Arg Pro Thr Gly Trp Lys Cys Phe Val Tyr His Phe Ala 
        115                 120                 125 

Val Phe Leu Ile Val Leu Val Cys Leu Ile Phe Ser Val Leu Ser Thr 
    130                 135                 140 

Ile Glu Gln Tyr Ala Ala Leu Ala Thr Gly Thr Leu Phe Trp Met Glu 
145                 150                 155                 160 

Ile Val Leu Val Val Phe Phe Gly Thr Glu Tyr Val Val Arg Leu Trp 
                165                 170                 175 

Ser Ala Gly Cys Arg Ser Lys Tyr Val Gly Leu Trp Gly Arg Leu Arg 
            180                 185                 190 

Phe Ala Arg Lys Pro Ile Ser Ile Ile Asp Leu Ile Val Val Val Ala 
        195                 200                 205 

Ser Met Val Val Leu Cys Val Gly Ser Lys Gly Gln Val Phe Ala Thr 
    210                 215                 220 

Ser Ala Ile Arg Gly Ile Arg Phe Leu Gln Ile Leu Arg Met Leu His 
225                 230                 235                 240 

Val Asp Arg Gln Gly Gly Thr Trp Arg Leu Leu Gly Ser Val Val Phe 
                245                 250                 255 

Ile His Arg Gln Glu Leu Ile Thr Thr Leu Tyr Ile Gly Phe Leu Gly 
            260                 265                 270 

Leu Ile Phe Ser Ser Tyr Phe Val Tyr Leu Ala Glu Lys Asp Ala Val 
        275                 280                 285 

Asn Glu Ser Gly Arg Val Glu Phe Gly Ser Tyr Ala Asp Ala Leu Trp 
    290                 295                 300 

Trp Gly Val Val Thr Val Thr Thr Ile Gly Tyr Gly Asp Lys Val Pro 
305                 310                 315                 320 

Gln Thr Trp Val Gly Lys Thr Ile Ala Ser Cys Phe Ser Val Phe Ala 
                325                 330                 335 

Ile Ser Phe Phe Ala Leu Pro Ala Gly Ile Leu Gly Ser Gly Phe Ala 
            340                 345                 350 

Leu Lys Val Gln Gln Lys Gln Arg Gln Lys His Phe Asn Arg Gln Ile 
        355                 360                 365 

Pro Ala Ala Ala Ser Leu Ile Gln Thr Ala Trp Arg Cys Tyr Ala Ala 
    370                 375                 380 

Glu Asn Pro Asp Ser Ser Thr Trp Lys Ile Tyr Ile Arg Lys Ala Pro 
385                 390                 395                 400 

Arg Ser His Thr Leu Leu Ser Pro Ser Pro Lys Pro Lys Lys Ser Val 
                405                 410                 415 

Val Val Lys Lys Lys Lys Phe Lys Leu Asp Lys Asp Asn Gly Val Thr 
            420                 425                 430 

Pro Gly Glu Lys Met Leu Thr Val Pro His Ile Thr Cys Asp Pro Pro 
        435                 440                 445 

Glu Glu Arg Arg Leu Asp His Phe Ser Val Asp Gly Tyr Asp Ser Ser 
    450                 455                 460 

Val Arg Lys Ser Pro Thr Leu Leu Glu Val Ser Met Pro His Phe Met 
465                 470                 475                 480 

Arg Thr Asn Ser Phe Ala Glu Asp Leu Asp Leu Glu Gly Glu Thr Leu 
                485                 490                 495 

Leu Thr Pro Ile Thr His Ile Ser Gln Leu Arg Glu His His Arg Ala 
            500                 505                 510 

Thr Ile Lys Val Ile Arg Arg Met Gln Tyr Phe Val Ala Lys Lys Lys 
        515                 520                 525 

Phe Gln Gln Ala Arg Lys Pro Tyr Asp Val Arg Asp Val Ile Glu Gln 
    530                 535                 540 

Tyr Ser Gln Gly His Leu Asn Leu Met Val Arg Ile Lys Glu Leu Gln 
545                 550                 555                 560 

Arg Arg Leu Asp Gln Ser Ile Gly Lys Pro Ser Leu Phe Ile Ser Val 
                565                 570                 575 

Ser Glu Lys Ser Lys Asp Arg Gly Ser Asn Thr Ile Gly Ala Arg Leu 
            580                 585                 590 

Asn Arg Val Glu Asp Lys Val Thr Gln Leu Asp Gln Arg Leu Ala Leu 
        595                 600                 605 

Ile Thr Asp Met Leu His Gln Leu Leu Ser Leu His Gly Gly Ser Thr 
    610                 615                 620 

Pro Gly Ser Gly Gly Pro Pro Arg Glu Gly Gly Ala His Ile Thr Gln 
625                 630                 635                 640 

Pro Cys Gly Ser Gly Gly Ser Val Asp Pro Glu Leu Phe Leu Pro Ser 
                645                 650                 655 

Asn Thr Leu Pro Thr Tyr Glu Gln Leu Thr Val Pro Arg Arg Gly Pro 
            660                 665                 670 

Asp Glu Gly Ser 
        675 

 
           
             3  
             6048  
             DNA  
             Homo sapiens  
             
               CDS  
               (1)..(6048)  
             
           
            3 

atg gca aac ttc cta tta cct cgg ggc acc agc agc ttc cgc agg ttc       48 
Met Ala Asn Phe Leu Leu Pro Arg Gly Thr Ser Ser Phe Arg Arg Phe 
  1               5                  10                  15 

aca cgg gag tcc ctg gca gcc atc gag aag cgc atg gcg gag aag caa       96 
Thr Arg Glu Ser Leu Ala Ala Ile Glu Lys Arg Met Ala Glu Lys Gln 
             20                  25                  30 

gcc cgc ggc tca acc acc ttg cag gag agc cga gag ggg ctg ccc gag      144 
Ala Arg Gly Ser Thr Thr Leu Gln Glu Ser Arg Glu Gly Leu Pro Glu 
         35                  40                  45 

gag gag gct ccc cgg ccc cag ctg gac ctg cag gcc tcc aaa aag ctg      192 
Glu Glu Ala Pro Arg Pro Gln Leu Asp Leu Gln Ala Ser Lys Lys Leu 
     50                  55                  60 

cca gat ctc tat ggc aat cca ccc caa gag ctc atc gga gag ccc ctg      240 
Pro Asp Leu Tyr Gly Asn Pro Pro Gln Glu Leu Ile Gly Glu Pro Leu 
 65                  70                  75                  80 

gag gac ctg gac ccc ttc tat agc acc caa aag act ttc atc gta ctg      288 
Glu Asp Leu Asp Pro Phe Tyr Ser Thr Gln Lys Thr Phe Ile Val Leu 
                 85                  90                  95 

aat aaa ggc aag acc atc ttc cgg ttc agt gcc acc aac gcc ttg tat      336 
Asn Lys Gly Lys Thr Ile Phe Arg Phe Ser Ala Thr Asn Ala Leu Tyr 
            100                 105                 110 

gtc ctc agt ccc ttc cac cca gtt cgg aga gcg gct gtg aag att ctg      384 
Val Leu Ser Pro Phe His Pro Val Arg Arg Ala Ala Val Lys Ile Leu 
        115                 120                 125 

gtt cac tcg ctc ttc aac atg ctc atc atg tgc acc atc ctc acc aac      432 
Val His Ser Leu Phe Asn Met Leu Ile Met Cys Thr Ile Leu Thr Asn 
    130                 135                 140 

tgc gtg ttc atg gcc cag cac gac cct cca ccc tgg acc aag tat gtc      480 
Cys Val Phe Met Ala Gln His Asp Pro Pro Pro Trp Thr Lys Tyr Val 
145                 150                 155                 160 

gag tac acc ttc acc gcc att tac acc ttt gag tct ctg gtc aag att      528 
Glu Tyr Thr Phe Thr Ala Ile Tyr Thr Phe Glu Ser Leu Val Lys Ile 
                165                 170                 175 

ctg gct cga gct ttc tgc ctg cac gcg ttc act ttc ctt cgg gac cca      576 
Leu Ala Arg Ala Phe Cys Leu His Ala Phe Thr Phe Leu Arg Asp Pro 
            180                 185                 190 

tgg aac tgg ctg gac ttt agt gtg att atc atg gca tac aca act gaa      624 
Trp Asn Trp Leu Asp Phe Ser Val Ile Ile Met Ala Tyr Thr Thr Glu 
        195                 200                 205 

ttt gtg gac ctg ggc aat gtc tca gcc tta cgc acc ttc cga gtc ctc      672 
Phe Val Asp Leu Gly Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu 
    210                 215                 220 

cgg gcc ctg aaa act ata tca gtc att tca ggg ctg aag acc atc gtg      720 
Arg Ala Leu Lys Thr Ile Ser Val Ile Ser Gly Leu Lys Thr Ile Val 
225                 230                 235                 240 

ggg gcc ctg atc cag tct gtg aag aag ctg gct gat gtg atg gtc ctc      768 
Gly Ala Leu Ile Gln Ser Val Lys Lys Leu Ala Asp Val Met Val Leu 
                245                 250                 255 

aca gtc ttc tgc ctc agc gtc ttt gcc ctc atc ggc ctg cag ctc ttc      816 
Thr Val Phe Cys Leu Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe 
            260                 265                 270 

atg ggc aac cta agg cac aag tgt gtg cgc aac ttc aca gcg ctc aac      864 
Met Gly Asn Leu Arg His Lys Cys Val Arg Asn Phe Thr Ala Leu Asn 
        275                 280                 285 

ggc acc aac ggc tcc gtg gag gcc gac ggc ttg gtc tgg gaa tcc ctg      912 
Gly Thr Asn Gly Ser Val Glu Ala Asp Gly Leu Val Trp Glu Ser Leu 
    290                 295                 300 

gac ctt tac ctc agt gat cca gaa aat tac ctg ctc aag aac ggc acc      960 
Asp Leu Tyr Leu Ser Asp Pro Glu Asn Tyr Leu Leu Lys Asn Gly Thr 
305                 310                 315                 320 

tct gat gtg tta ctg tgt ggg aac agc tct gac gct ggg aca tgt ccg     1008 
Ser Asp Val Leu Leu Cys Gly Asn Ser Ser Asp Ala Gly Thr Cys Pro 
                325                 330                 335 

gag ggc tac cgg tgc cta aag gca ggc gag aac ccc gac cac ggc tac     1056 
Glu Gly Tyr Arg Cys Leu Lys Ala Gly Glu Asn Pro Asp His Gly Tyr 
            340                 345                 350 

acc agc ttc gat tcc ttt gcc tgg gcc ttt ctt gca ctc ttc cgc ctg     1104 
Thr Ser Phe Asp Ser Phe Ala Trp Ala Phe Leu Ala Leu Phe Arg Leu 
        355                 360                 365 

atg acg cag gac tgc tgg gag cgc ctc tat cag cag acc ctc agg tcc     1152 
Met Thr Gln Asp Cys Trp Glu Arg Leu Tyr Gln Gln Thr Leu Arg Ser 
    370                 375                 380 

gca ggg aag atc tac atg atc ttc ttc atg ctt gtc atc ttc ctg ggg     1200 
Ala Gly Lys Ile Tyr Met Ile Phe Phe Met Leu Val Ile Phe Leu Gly 
385                 390                 395                 400 

tcc ttc tac ctg gtg aac ctg atc ctg gcc gtg gtc gca atg gcc tat     1248 
Ser Phe Tyr Leu Val Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr 
                405                 410                 415 

gag gag caa aac caa gcc acc atc gct gag acc gag gag aag gaa aag     1296 
Glu Glu Gln Asn Gln Ala Thr Ile Ala Glu Thr Glu Glu Lys Glu Lys 
            420                 425                 430 

cgc ttc cag gag gcc atg gaa atg ctc aag aaa gaa cac gag gcc ctc     1344 
Arg Phe Gln Glu Ala Met Glu Met Leu Lys Lys Glu His Glu Ala Leu 
        435                 440                 445 

acc atc agg ggt gtg gat acc gtg tcc cgt agc tcc ttg gag atg tcc     1392 
Thr Ile Arg Gly Val Asp Thr Val Ser Arg Ser Ser Leu Glu Met Ser 
    450                 455                 460 

cct ttg gcc cca gta aac agc cat gag aga aga agc aag agg aga aaa     1440 
Pro Leu Ala Pro Val Asn Ser His Glu Arg Arg Ser Lys Arg Arg Lys 
465                 470                 475                 480 

cgg atg tct tca gga act gag gag tgt ggg gag gac agg ctc ccc aag     1488 
Arg Met Ser Ser Gly Thr Glu Glu Cys Gly Glu Asp Arg Leu Pro Lys 
                485                 490                 495 

tct gac tca gaa gat ggt ccc aga gca atg aat cat ctc agc ctc acc     1536 
Ser Asp Ser Glu Asp Gly Pro Arg Ala Met Asn His Leu Ser Leu Thr 
            500                 505                 510 

cgt ggc ctc agc agg act tct atg aag cca cgt tcc agc cgc ggg agc     1584 
Arg Gly Leu Ser Arg Thr Ser Met Lys Pro Arg Ser Ser Arg Gly Ser 
        515                 520                 525 

att ttc acc ttt cgc agg cga gac ctg ggt tct gaa gca gat ttt gca     1632 
Ile Phe Thr Phe Arg Arg Arg Asp Leu Gly Ser Glu Ala Asp Phe Ala 
    530                 535                 540 

gat gat gaa aac agc aca gcg cgg gag agc gag agc cac cac aca tca     1680 
Asp Asp Glu Asn Ser Thr Ala Arg Glu Ser Glu Ser His His Thr Ser 
545                 550                 555                 560 

ctg ctg gtg ccc tgg ccc ctg cgc cgg acc agt gcc cag gga cag ccc     1728 
Leu Leu Val Pro Trp Pro Leu Arg Arg Thr Ser Ala Gln Gly Gln Pro 
                565                 570                 575 

agt ccc gga acc tcg gct cct ggc cac gcc ctc cat ggc aaa aag aac     1776 
Ser Pro Gly Thr Ser Ala Pro Gly His Ala Leu His Gly Lys Lys Asn 
            580                 585                 590 

agc act gtg gac tgc aat ggg gtg gtc tca tta ctg ggg gca ggc gac     1824 
Ser Thr Val Asp Cys Asn Gly Val Val Ser Leu Leu Gly Ala Gly Asp 
        595                 600                 605 

cca gag gcc aca tcc cca gga agc cac ctc ctc cgc cct gtg atg cta     1872 
Pro Glu Ala Thr Ser Pro Gly Ser His Leu Leu Arg Pro Val Met Leu 
    610                 615                 620 

gag cac ccg cca gac acg acc acg cca tcg gag gag cca ggc ggc ccc     1920 
Glu His Pro Pro Asp Thr Thr Thr Pro Ser Glu Glu Pro Gly Gly Pro 
625                 630                 635                 640 

cag atg ctg acc tcc cag gct ccg tgt gta gat ggc ttc gag gag cca     1968 
Gln Met Leu Thr Ser Gln Ala Pro Cys Val Asp Gly Phe Glu Glu Pro 
                645                 650                 655 

gga gca cgg cag cgg gcc ctc agc gca gtc agc gtc ctc aca agc gca     2016 
Gly Ala Arg Gln Arg Ala Leu Ser Ala Val Ser Val Leu Thr Ser Ala 
            660                 665                 670 

ctg gaa gag tta gag gag tct cgc cac aag tgt cca cca tgc tgg aac     2064 
Leu Glu Glu Leu Glu Glu Ser Arg His Lys Cys Pro Pro Cys Trp Asn 
        675                 680                 685 

cgt ctc gcc cag cgc tac ctg atc tgg gag tgc tgc ccg ctg tgg atg     2112 
Arg Leu Ala Gln Arg Tyr Leu Ile Trp Glu Cys Cys Pro Leu Trp Met 
    690                 695                 700 

tcc atc aag cag gga gtg aag ttg gtg gtc atg gac ccg ttt act gac     2160 
Ser Ile Lys Gln Gly Val Lys Leu Val Val Met Asp Pro Phe Thr Asp 
705                 710                 715                 720 

ctc acc atc act atg tgc atc gta ctc aac aca ctc ttc atg gcg ctg     2208 
Leu Thr Ile Thr Met Cys Ile Val Leu Asn Thr Leu Phe Met Ala Leu 
                725                 730                 735 

gag cac tac aac atg aca agt gaa ttc gag gag atg ctg cag gtc gga     2256 
Glu His Tyr Asn Met Thr Ser Glu Phe Glu Glu Met Leu Gln Val Gly 
            740                 745                 750 

aac ctg gtc ttc aca ggg att ttc aca gca gag atg acc ttc aag atc     2304 
Asn Leu Val Phe Thr Gly Ile Phe Thr Ala Glu Met Thr Phe Lys Ile 
        755                 760                 765 

att gcc ctc gac ccc tac tac tac ttc caa cag ggc tgg aac atc ttc     2352 
Ile Ala Leu Asp Pro Tyr Tyr Tyr Phe Gln Gln Gly Trp Asn Ile Phe 
    770                 775                 780 

gac agc atc atc gtc atc ctt agc ctc atg gag ctg ggc ctg tcc cgc     2400 
Asp Ser Ile Ile Val Ile Leu Ser Leu Met Glu Leu Gly Leu Ser Arg 
785                 790                 795                 800 

atg agc aac ttg tcg gtg ctg cgc tcc ttc cgc ctg ctg cgg gtc ttc     2448 
Met Ser Asn Leu Ser Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe 
                805                 810                 815 

aag ctg gcc aaa tca tgg ccc acc ctg aac aca ctc atc aag atc atc     2496 
Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn Thr Leu Ile Lys Ile Ile 
            820                 825                 830 

ggg aac tca gtg ggg gca ctg ggg aac ctg aca ctg gtg cta gcc atc     2544 
Gly Asn Ser Val Gly Ala Leu Gly Asn Leu Thr Leu Val Leu Ala Ile 
        835                 840                 845 

atc gtg ttc atc ttt gct gtg gtg ggc atg cag ctc ttt ggc aag aac     2592 
Ile Val Phe Ile Phe Ala Val Val Gly Met Gln Leu Phe Gly Lys Asn 
    850                 855                 860 

tac tcg gag ctg agg gac agc gac tca ggc ctg ctg cct cgc tgg cac     2640 
Tyr Ser Glu Leu Arg Asp Ser Asp Ser Gly Leu Leu Pro Arg Trp His 
865                 870                 875                 880 

atg atg gac ttc ttt cat gcc ttc cta atc atc ttc cgc atc ctc tgt     2688 
Met Met Asp Phe Phe His Ala Phe Leu Ile Ile Phe Arg Ile Leu Cys 
                885                 890                 895 

gga gag tgg atc gag acc atg tgg gac tgc atg gag gtg tcg ggg cag     2736 
Gly Glu Trp Ile Glu Thr Met Trp Asp Cys Met Glu Val Ser Gly Gln 
            900                 905                 910 

tca tta tgc ctg ctg gtc ttc ttg ctt gtt atg gtc att ggc aac ctt     2784 
Ser Leu Cys Leu Leu Val Phe Leu Leu Val Met Val Ile Gly Asn Leu 
        915                 920                 925 

gtg gtc ctg aat ctc ttc ctg gcc ttg ctg ctc agc tcc ttc agt gca     2832 
Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe Ser Ala 
    930                 935                 940 

gac aac ctc aca gcc cct gat gag gac aga gag atg aac aac ctc cag     2880 
Asp Asn Leu Thr Ala Pro Asp Glu Asp Arg Glu Met Asn Asn Leu Gln 
945                 950                 955                 960 

ctg gcc ctg gcc cgc atc cag agg ggc ctg cgc ttt gtc aag cgg acc     2928 
Leu Ala Leu Ala Arg Ile Gln Arg Gly Leu Arg Phe Val Lys Arg Thr 
                965                 970                 975 

acc tgg gat ttc tgc tgt ggt ctc ctg cgg cac cgg cct cag aag ccc     2976 
Thr Trp Asp Phe Cys Cys Gly Leu Leu Arg His Arg Pro Gln Lys Pro 
            980                 985                 990 

gca gcc ctt gcc gcc cag ggc cag ctg ccc agc tgc att gcc acc ccc     3024 
Ala Ala Leu Ala Ala Gln Gly Gln Leu Pro Ser Cys Ile Ala Thr Pro 
        995                 1000                1005 

tac tcc ccg cca ccc cca gag acg gag aag gtg cct ccc acc cgc aag     3072 
Tyr Ser Pro Pro Pro Pro Glu Thr Glu Lys Val Pro Pro Thr Arg Lys 
    1010                1015                1020 

gaa aca cag ttt gag gaa ggc gag caa cca ggc cag ggc acc ccc ggg     3120 
Glu Thr Gln Phe Glu Glu Gly Glu Gln Pro Gly Gln Gly Thr Pro Gly 
1025                1030                1035                1040 

gat cca gag ccc gtg tgt gtg ccc atc gct gtg gcc gag tca gac aca     3168 
Asp Pro Glu Pro Val Cys Val Pro Ile Ala Val Ala Glu Ser Asp Thr 
                1045                1050                1055 

gat gac caa gaa gag gat gag gag aac agc ctg ggc acg gag gag gag     3216 
Asp Asp Gln Glu Glu Asp Glu Glu Asn Ser Leu Gly Thr Glu Glu Glu 
            1060                1065                1070 

tcc agc aag cag cag gaa tcc cag cct gtg tcc ggc tgg ccc aga ggc     3264 
Ser Ser Lys Gln Gln Glu Ser Gln Pro Val Ser Gly Trp Pro Arg Gly 
        1075                1080                1085 

cct ccg gat tcc agg acc tgg agc cag gtg tca gcg act gcc tcc tct     3312 
Pro Pro Asp Ser Arg Thr Trp Ser Gln Val Ser Ala Thr Ala Ser Ser 
    1090                1095                1100 

gag gcc gag gcc agt gca tct cag gcc gac tgg cgg cag cag tgg aaa     3360 
Glu Ala Glu Ala Ser Ala Ser Gln Ala Asp Trp Arg Gln Gln Trp Lys 
1105                1110                1115                1120 

gcg gaa ccc cag gcc cca ggg tgc ggt gag acc cca gag gac agt tgc     3408 
Ala Glu Pro Gln Ala Pro Gly Cys Gly Glu Thr Pro Glu Asp Ser Cys 
                1125                1130                1135 

tcc gag ggc agc aca gca gac atg acc aac acc gct gag ctc ctg gag     3456 
Ser Glu Gly Ser Thr Ala Asp Met Thr Asn Thr Ala Glu Leu Leu Glu 
            1140                1145                1150 

cag atc cct gac ctc ggc cag gat gtc aag gac cca gag gac tgc ttc     3504 
Gln Ile Pro Asp Leu Gly Gln Asp Val Lys Asp Pro Glu Asp Cys Phe 
        1155                1160                1165 

act gaa ggc tgt gtc cgg cgc tgt ccc tgc tgt gcg gtg gac acc aca     3552 
Thr Glu Gly Cys Val Arg Arg Cys Pro Cys Cys Ala Val Asp Thr Thr 
    1170                1175                1180 

cag gcc cca ggg aag gtc tgg tgg cgg ttg cgc aag acc tgc tac cac     3600 
Gln Ala Pro Gly Lys Val Trp Trp Arg Leu Arg Lys Thr Cys Tyr His 
1185                1190                1195                1200 

atc gtg gag cac agc tgg ttc gag aca ttc atc atc ttc atg atc cta     3648 
Ile Val Glu His Ser Trp Phe Glu Thr Phe Ile Ile Phe Met Ile Leu 
                1205                1210                1215 

ctc agc agt gga gcg ctg gcc ttc gag gac atc tac cta gag gag cgg     3696 
Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Leu Glu Glu Arg 
            1220                1225                1230 

aag acc atc aag gtt ctg ctt gag tat gcc gac aag atg ttc aca tat     3744 
Lys Thr Ile Lys Val Leu Leu Glu Tyr Ala Asp Lys Met Phe Thr Tyr 
        1235                1240                1245 

gtc ttc gtg ctg gag atg ctg ctc aag tgg gtg gcc tac ggc ttc aag     3792 
Val Phe Val Leu Glu Met Leu Leu Lys Trp Val Ala Tyr Gly Phe Lys 
    1250                1255                1260 

aag tac ttc acc aat gcc tgg tgc tgg ctc gac ttc ctc atc gta gac     3840 
Lys Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val Asp 
1265                1270                1275                1280 

gtc tct ctg gtc agc ctg gtg gcc aac acc ctg ggc ttt gcc gag atg     3888 
Val Ser Leu Val Ser Leu Val Ala Asn Thr Leu Gly Phe Ala Glu Met 
                1285                1290                1295 

ggc ccc atc aag tca ctg cgg acg ctg cgt gca ctc cgt cct ctg aga     3936 
Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg Pro Leu Arg 
            1300                1305                1310 

gct ctg tca cga ttt gag ggc atg agg gtg gtg gtc aat gcc ctg gtg     3984 
Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val Asn Ala Leu Val 
        1315                1320                1325 

ggc gcc atc ccg tcc atc atg aac gtc ctc ctc gtc tgc ctc atc ttc     4032 
Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu Val Cys Leu Ile Phe 
    1330                1335                1340 

tgg ctc atc ttc agc atc atg ggc gtg aac ctc ttt gcg ggg aag ttt     4080 
Trp Leu Ile Phe Ser Ile Met Gly Val Asn Leu Phe Ala Gly Lys Phe 
1345                1350                1355                1360 

ggg agg tgc atc aac cag aca gag gga gac ttg cct ttg aac tac acc     4128 
Gly Arg Cys Ile Asn Gln Thr Glu Gly Asp Leu Pro Leu Asn Tyr Thr 
                1365                1370                1375 

atc gtg aac aac aag agc cag tgt gag tcc ttg aac ttg acc gga gaa     4176 
Ile Val Asn Asn Lys Ser Gln Cys Glu Ser Leu Asn Leu Thr Gly Glu 
            1380                1385                1390 

ttg tac tgg acc aag gtg aaa gtc aac ttt gac aac gtg ggg gcc ggg     4224 
Leu Tyr Trp Thr Lys Val Lys Val Asn Phe Asp Asn Val Gly Ala Gly 
        1395                1400                1405 

tac ctg gcc ctt ctg cag gtg gca aca ttt aaa ggc tgg atg gac att     4272 
Tyr Leu Ala Leu Leu Gln Val Ala Thr Phe Lys Gly Trp Met Asp Ile 
    1410                1415                1420 

atg tat gca gct gtg gac tcc agg ggg tat gaa gag cag cct cag tgg     4320 
Met Tyr Ala Ala Val Asp Ser Arg Gly Tyr Glu Glu Gln Pro Gln Trp 
1425                1430                1435                1440 

gaa tac aac ctc tac atg tac atc tat ttt gtc att ttc atc atc ttt     4368 
Glu Tyr Asn Leu Tyr Met Tyr Ile Tyr Phe Val Ile Phe Ile Ile Phe 
                1445                1450                1455 

ggg tct ttc ttc acc ctg aac ctc ttt att ggt gtc atc att gac aac     4416 
Gly Ser Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn 
            1460                1465                1470 

ttc aac caa cag aag aaa aag tta ggg ggc cag gac atc ttc atg aca     4464 
Phe Asn Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr 
        1475                1480                1485 

gag gag cag aag aag tac tac aat gcc atg aag aag ctg ggc tcc aag     4512 
Glu Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys 
    1490                1495                1500 

aag ccc cag aag ccc atc cca cgg ccc ctg aac aag tac cag ggc ttc     4560 
Lys Pro Gln Lys Pro Ile Pro Arg Pro Leu Asn Lys Tyr Gln Gly Phe 
1505                1510                1515                1520 

ata ttc gac att gtg acc aag cag gcc ttt gac gtc acc atc atg ttt     4608 
Ile Phe Asp Ile Val Thr Lys Gln Ala Phe Asp Val Thr Ile Met Phe 
                1525                1530                1535 

ctg atc tgc ttg aat atg gtg acc atg atg gtg gag aca gat gac caa     4656 
Leu Ile Cys Leu Asn Met Val Thr Met Met Val Glu Thr Asp Asp Gln 
            1540                1545                1550 

agt cct gag aaa atc aac atc ttg gcc aag atc aac ctg ctc ttt gtg     4704 
Ser Pro Glu Lys Ile Asn Ile Leu Ala Lys Ile Asn Leu Leu Phe Val 
        1555                1560                1565 

gcc atc ttc aca ggc gag tgt att gtc aag ctg gct gcc ctg cgc cac     4752 
Ala Ile Phe Thr Gly Glu Cys Ile Val Lys Leu Ala Ala Leu Arg His 
    1570                1575                1580 

tac tac ttc acc aac agc tgg aat atc ttc gac ttc gtg gtt gtc atc     4800 
Tyr Tyr Phe Thr Asn Ser Trp Asn Ile Phe Asp Phe Val Val Val Ile 
1585                1590                1595                1600 

ctc tcc atc gtg ggc act gtg ctc tcg gac atc atc cag aag tac ttc     4848 
Leu Ser Ile Val Gly Thr Val Leu Ser Asp Ile Ile Gln Lys Tyr Phe 
                1605                1610                1615 

ttc tcc ccg acg ctc ttc cga gtc atc cgc ctg gcc cga ata ggc cgc     4896 
Phe Ser Pro Thr Leu Phe Arg Val Ile Arg Leu Ala Arg Ile Gly Arg 
            1620                1625                1630 

atc ctc aga ctg atc cga ggg gcc aag ggg atc cgc acg ctg ctc ttt     4944 
Ile Leu Arg Leu Ile Arg Gly Ala Lys Gly Ile Arg Thr Leu Leu Phe 
        1635                1640                1645 

gcc ctc atg atg tcc ctg cct gcc ctc ttc aac atc ggg ctg ctg ctc     4992 
Ala Leu Met Met Ser Leu Pro Ala Leu Phe Asn Ile Gly Leu Leu Leu 
    1650                1655                1660 

ttc ctc gtc atg ttc atc tac tcc atc ttt ggc atg gcc aac ttc gct     5040 
Phe Leu Val Met Phe Ile Tyr Ser Ile Phe Gly Met Ala Asn Phe Ala 
1665                1670                1675                1680 

tat gtc aag tgg gag gct ggc atc gac gac atg ttc aac ttc cag acc     5088 
Tyr Val Lys Trp Glu Ala Gly Ile Asp Asp Met Phe Asn Phe Gln Thr 
                1685                1690                1695 

ttc gcc aac agc atg ctg tgc ctc ttc cag atc acc acg tcg gcc ggc     5136 
Phe Ala Asn Ser Met Leu Cys Leu Phe Gln Ile Thr Thr Ser Ala Gly 
            1700                1705                1710 

tgg gat ggc ctc ctc agc ccc atc ctc aac act ggg ccg ccc tac tgc     5184 
Trp Asp Gly Leu Leu Ser Pro Ile Leu Asn Thr Gly Pro Pro Tyr Cys 
        1715                1720                1725 

gac ccc act ctg ccc aac agc aat ggc tct cgg ggg gac tgc ggg agc     5232 
Asp Pro Thr Leu Pro Asn Ser Asn Gly Ser Arg Gly Asp Cys Gly Ser 
    1730                1735                1740 

cca gcc gtg ggc atc ctc ttc ttc acc acc tac atc atc atc tcc ttc     5280 
Pro Ala Val Gly Ile Leu Phe Phe Thr Thr Tyr Ile Ile Ile Ser Phe 
1745                1750                1755                1760 

ctc atc gtg gtc aac atg tac att gcc atc atc ctg gag aac ttc agc     5328 
Leu Ile Val Val Asn Met Tyr Ile Ala Ile Ile Leu Glu Asn Phe Ser 
                1765                1770                1775 

gtg gcc acg gag gag agc acc gag ccc ctg agt gag gac gac ttc gat     5376 
Val Ala Thr Glu Glu Ser Thr Glu Pro Leu Ser Glu Asp Asp Phe Asp 
            1780                1785                1790 

atg ttc tat gag atc tgg gag aaa ttt gac cca gag gcc act cag ttt     5424 
Met Phe Tyr Glu Ile Trp Glu Lys Phe Asp Pro Glu Ala Thr Gln Phe 
        1795                1800                1805 

att gag tat tcg gtc ctg tct gac ttt gcc gac gcc ctg tct gag cca     5472 
Ile Glu Tyr Ser Val Leu Ser Asp Phe Ala Asp Ala Leu Ser Glu Pro 
    1810                1815                1820 

ctc cgt atc gcc aag ccc aac cag ata agc ctc atc aac atg gac ctg     5520 
Leu Arg Ile Ala Lys Pro Asn Gln Ile Ser Leu Ile Asn Met Asp Leu 
1825                1830                1835                1840 

ccc atg gtg agt ggg gac cgc atc cat tgc atg gac att ctc ttt gcc     5568 
Pro Met Val Ser Gly Asp Arg Ile His Cys Met Asp Ile Leu Phe Ala 
                1845                1850                1855 

ttc acc aaa agg gtc ctg ggg gag tct ggg gag atg gac gcc ctg aag     5616 
Phe Thr Lys Arg Val Leu Gly Glu Ser Gly Glu Met Asp Ala Leu Lys 
            1860                1865                1870 

atc cag atg gag gag aag ttc atg gca gcc aac cca tcc aag atc tcc     5664 
Ile Gln Met Glu Glu Lys Phe Met Ala Ala Asn Pro Ser Lys Ile Ser 
        1875                1880                1885 

tac gag ccc atc acc acc aca ctc cgg cgc aag cac gaa gag gtg tcg     5712 
Tyr Glu Pro Ile Thr Thr Thr Leu Arg Arg Lys His Glu Glu Val Ser 
    1890                1895                1900 

gcc atg gtt atc cag aga gcc ttc cgc agg cac ctg ctg caa cgc tct     5760 
Ala Met Val Ile Gln Arg Ala Phe Arg Arg His Leu Leu Gln Arg Ser 
1905                1910                1915                1920 

ttg aag cat gcc tcc ttc ctc ttc cgt cag cag gcg ggc agc ggc ctc     5808 
Leu Lys His Ala Ser Phe Leu Phe Arg Gln Gln Ala Gly Ser Gly Leu 
                1925                1930                1935 

tcc gaa gag gat gcc cct gag cga gag ggc ctc atc gcc tac gtg atg     5856 
Ser Glu Glu Asp Ala Pro Glu Arg Glu Gly Leu Ile Ala Tyr Val Met 
            1940                1945                1950 

agt gag aac ttc tcc cga ccc ctt ggc cca ccc tcc agc tcc tcc atc     5904 
Ser Glu Asn Phe Ser Arg Pro Leu Gly Pro Pro Ser Ser Ser Ser Ile 
        1955                1960                1965 

tcc tcc act tcc ttc cca ccc tcc tat gac agt gtc act aga gcc acc     5952 
Ser Ser Thr Ser Phe Pro Pro Ser Tyr Asp Ser Val Thr Arg Ala Thr 
    1970                1975                1980 

agc gat aac ctc cag gtg cgg ggg tct gac tac agc cac agt gaa gat     6000 
Ser Asp Asn Leu Gln Val Arg Gly Ser Asp Tyr Ser His Ser Glu Asp 
1985                1990                1995                2000 

ctc gcc gac ttc ccc cct tct ccg gac agg gac cgt gag tcc atc gtg     6048 
Leu Ala Asp Phe Pro Pro Ser Pro Asp Arg Asp Arg Glu Ser Ile Val 
                2005                2010                2015 

 
           
             4  
             2016  
             PRT  
             Homo sapiens  
           
            4 

Met Ala Asn Phe Leu Leu Pro Arg Gly Thr Ser Ser Phe Arg Arg Phe 
  1               5                  10                  15 

Thr Arg Glu Ser Leu Ala Ala Ile Glu Lys Arg Met Ala Glu Lys Gln 
             20                  25                  30 

Ala Arg Gly Ser Thr Thr Leu Gln Glu Ser Arg Glu Gly Leu Pro Glu 
         35                  40                  45 

Glu Glu Ala Pro Arg Pro Gln Leu Asp Leu Gln Ala Ser Lys Lys Leu 
     50                  55                  60 

Pro Asp Leu Tyr Gly Asn Pro Pro Gln Glu Leu Ile Gly Glu Pro Leu 
 65                  70                  75                  80 

Glu Asp Leu Asp Pro Phe Tyr Ser Thr Gln Lys Thr Phe Ile Val Leu 
                 85                  90                  95 

Asn Lys Gly Lys Thr Ile Phe Arg Phe Ser Ala Thr Asn Ala Leu Tyr 
            100                 105                 110 

Val Leu Ser Pro Phe His Pro Val Arg Arg Ala Ala Val Lys Ile Leu 
        115                 120                 125 

Val His Ser Leu Phe Asn Met Leu Ile Met Cys Thr Ile Leu Thr Asn 
    130                 135                 140 

Cys Val Phe Met Ala Gln His Asp Pro Pro Pro Trp Thr Lys Tyr Val 
145                 150                 155                 160 

Glu Tyr Thr Phe Thr Ala Ile Tyr Thr Phe Glu Ser Leu Val Lys Ile 
                165                 170                 175 

Leu Ala Arg Ala Phe Cys Leu His Ala Phe Thr Phe Leu Arg Asp Pro 
            180                 185                 190 

Trp Asn Trp Leu Asp Phe Ser Val Ile Ile Met Ala Tyr Thr Thr Glu 
        195                 200                 205 

Phe Val Asp Leu Gly Asn Val Ser Ala Leu Arg Thr Phe Arg Val Leu 
    210                 215                 220 

Arg Ala Leu Lys Thr Ile Ser Val Ile Ser Gly Leu Lys Thr Ile Val 
225                 230                 235                 240 

Gly Ala Leu Ile Gln Ser Val Lys Lys Leu Ala Asp Val Met Val Leu 
                245                 250                 255 

Thr Val Phe Cys Leu Ser Val Phe Ala Leu Ile Gly Leu Gln Leu Phe 
            260                 265                 270 

Met Gly Asn Leu Arg His Lys Cys Val Arg Asn Phe Thr Ala Leu Asn 
        275                 280                 285 

Gly Thr Asn Gly Ser Val Glu Ala Asp Gly Leu Val Trp Glu Ser Leu 
    290                 295                 300 

Asp Leu Tyr Leu Ser Asp Pro Glu Asn Tyr Leu Leu Lys Asn Gly Thr 
305                 310                 315                 320 

Ser Asp Val Leu Leu Cys Gly Asn Ser Ser Asp Ala Gly Thr Cys Pro 
                325                 330                 335 

Glu Gly Tyr Arg Cys Leu Lys Ala Gly Glu Asn Pro Asp His Gly Tyr 
            340                 345                 350 

Thr Ser Phe Asp Ser Phe Ala Trp Ala Phe Leu Ala Leu Phe Arg Leu 
        355                 360                 365 

Met Thr Gln Asp Cys Trp Glu Arg Leu Tyr Gln Gln Thr Leu Arg Ser 
    370                 375                 380 

Ala Gly Lys Ile Tyr Met Ile Phe Phe Met Leu Val Ile Phe Leu Gly 
385                 390                 395                 400 

Ser Phe Tyr Leu Val Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr 
                405                 410                 415 

Glu Glu Gln Asn Gln Ala Thr Ile Ala Glu Thr Glu Glu Lys Glu Lys 
            420                 425                 430 

Arg Phe Gln Glu Ala Met Glu Met Leu Lys Lys Glu His Glu Ala Leu 
        435                 440                 445 

Thr Ile Arg Gly Val Asp Thr Val Ser Arg Ser Ser Leu Glu Met Ser 
    450                 455                 460 

Pro Leu Ala Pro Val Asn Ser His Glu Arg Arg Ser Lys Arg Arg Lys 
465                 470                 475                 480 

Arg Met Ser Ser Gly Thr Glu Glu Cys Gly Glu Asp Arg Leu Pro Lys 
                485                 490                 495 

Ser Asp Ser Glu Asp Gly Pro Arg Ala Met Asn His Leu Ser Leu Thr 
            500                 505                 510 

Arg Gly Leu Ser Arg Thr Ser Met Lys Pro Arg Ser Ser Arg Gly Ser 
        515                 520                 525 

Ile Phe Thr Phe Arg Arg Arg Asp Leu Gly Ser Glu Ala Asp Phe Ala 
    530                 535                 540 

Asp Asp Glu Asn Ser Thr Ala Arg Glu Ser Glu Ser His His Thr Ser 
545                 550                 555                 560 

Leu Leu Val Pro Trp Pro Leu Arg Arg Thr Ser Ala Gln Gly Gln Pro 
                565                 570                 575 

Ser Pro Gly Thr Ser Ala Pro Gly His Ala Leu His Gly Lys Lys Asn 
            580                 585                 590 

Ser Thr Val Asp Cys Asn Gly Val Val Ser Leu Leu Gly Ala Gly Asp 
        595                 600                 605 

Pro Glu Ala Thr Ser Pro Gly Ser His Leu Leu Arg Pro Val Met Leu 
    610                 615                 620 

Glu His Pro Pro Asp Thr Thr Thr Pro Ser Glu Glu Pro Gly Gly Pro 
625                 630                 635                 640 

Gln Met Leu Thr Ser Gln Ala Pro Cys Val Asp Gly Phe Glu Glu Pro 
                645                 650                 655 

Gly Ala Arg Gln Arg Ala Leu Ser Ala Val Ser Val Leu Thr Ser Ala 
            660                 665                 670 

Leu Glu Glu Leu Glu Glu Ser Arg His Lys Cys Pro Pro Cys Trp Asn 
        675                 680                 685 

Arg Leu Ala Gln Arg Tyr Leu Ile Trp Glu Cys Cys Pro Leu Trp Met 
    690                 695                 700 

Ser Ile Lys Gln Gly Val Lys Leu Val Val Met Asp Pro Phe Thr Asp 
705                 710                 715                 720 

Leu Thr Ile Thr Met Cys Ile Val Leu Asn Thr Leu Phe Met Ala Leu 
                725                 730                 735 

Glu His Tyr Asn Met Thr Ser Glu Phe Glu Glu Met Leu Gln Val Gly 
            740                 745                 750 

Asn Leu Val Phe Thr Gly Ile Phe Thr Ala Glu Met Thr Phe Lys Ile 
        755                 760                 765 

Ile Ala Leu Asp Pro Tyr Tyr Tyr Phe Gln Gln Gly Trp Asn Ile Phe 
    770                 775                 780 

Asp Ser Ile Ile Val Ile Leu Ser Leu Met Glu Leu Gly Leu Ser Arg 
785                 790                 795                 800 

Met Ser Asn Leu Ser Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe 
                805                 810                 815 

Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn Thr Leu Ile Lys Ile Ile 
            820                 825                 830 

Gly Asn Ser Val Gly Ala Leu Gly Asn Leu Thr Leu Val Leu Ala Ile 
        835                 840                 845 

Ile Val Phe Ile Phe Ala Val Val Gly Met Gln Leu Phe Gly Lys Asn 
    850                 855                 860 

Tyr Ser Glu Leu Arg Asp Ser Asp Ser Gly Leu Leu Pro Arg Trp His 
865                 870                 875                 880 

Met Met Asp Phe Phe His Ala Phe Leu Ile Ile Phe Arg Ile Leu Cys 
                885                 890                 895 

Gly Glu Trp Ile Glu Thr Met Trp Asp Cys Met Glu Val Ser Gly Gln 
            900                 905                 910 

Ser Leu Cys Leu Leu Val Phe Leu Leu Val Met Val Ile Gly Asn Leu 
        915                 920                 925 

Val Val Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Ser Phe Ser Ala 
    930                 935                 940 

Asp Asn Leu Thr Ala Pro Asp Glu Asp Arg Glu Met Asn Asn Leu Gln 
945                 950                 955                 960 

Leu Ala Leu Ala Arg Ile Gln Arg Gly Leu Arg Phe Val Lys Arg Thr 
                965                 970                 975 

Thr Trp Asp Phe Cys Cys Gly Leu Leu Arg His Arg Pro Gln Lys Pro 
            980                 985                 990 

Ala Ala Leu Ala Ala Gln Gly Gln Leu Pro Ser Cys Ile Ala Thr Pro 
        995                 1000                1005 

Tyr Ser Pro Pro Pro Pro Glu Thr Glu Lys Val Pro Pro Thr Arg Lys 
    1010                1015                1020 

Glu Thr Gln Phe Glu Glu Gly Glu Gln Pro Gly Gln Gly Thr Pro Gly 
1025                1030                1035                1040 

Asp Pro Glu Pro Val Cys Val Pro Ile Ala Val Ala Glu Ser Asp Thr 
                1045                1050                1055 

Asp Asp Gln Glu Glu Asp Glu Glu Asn Ser Leu Gly Thr Glu Glu Glu 
            1060                1065                1070 

Ser Ser Lys Gln Gln Glu Ser Gln Pro Val Ser Gly Trp Pro Arg Gly 
        1075                1080                1085 

Pro Pro Asp Ser Arg Thr Trp Ser Gln Val Ser Ala Thr Ala Ser Ser 
    1090                1095                1100 

Glu Ala Glu Ala Ser Ala Ser Gln Ala Asp Trp Arg Gln Gln Trp Lys 
1105                1110                1115                1120 

Ala Glu Pro Gln Ala Pro Gly Cys Gly Glu Thr Pro Glu Asp Ser Cys 
                1125                1130                1135 

Ser Glu Gly Ser Thr Ala Asp Met Thr Asn Thr Ala Glu Leu Leu Glu 
            1140                1145                1150 

Gln Ile Pro Asp Leu Gly Gln Asp Val Lys Asp Pro Glu Asp Cys Phe 
        1155                1160                1165 

Thr Glu Gly Cys Val Arg Arg Cys Pro Cys Cys Ala Val Asp Thr Thr 
    1170                1175                1180 

Gln Ala Pro Gly Lys Val Trp Trp Arg Leu Arg Lys Thr Cys Tyr His 
1185                1190                1195                1200 

Ile Val Glu His Ser Trp Phe Glu Thr Phe Ile Ile Phe Met Ile Leu 
                1205                1210                1215 

Leu Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Leu Glu Glu Arg 
            1220                1225                1230 

Lys Thr Ile Lys Val Leu Leu Glu Tyr Ala Asp Lys Met Phe Thr Tyr 
        1235                1240                1245 

Val Phe Val Leu Glu Met Leu Leu Lys Trp Val Ala Tyr Gly Phe Lys 
    1250                1255                1260 

Lys Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val Asp 
1265                1270                1275                1280 

Val Ser Leu Val Ser Leu Val Ala Asn Thr Leu Gly Phe Ala Glu Met 
                1285                1290                1295 

Gly Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg Pro Leu Arg 
            1300                1305                1310 

Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val Asn Ala Leu Val 
        1315                1320                1325 

Gly Ala Ile Pro Ser Ile Met Asn Val Leu Leu Val Cys Leu Ile Phe 
    1330                1335                1340 

Trp Leu Ile Phe Ser Ile Met Gly Val Asn Leu Phe Ala Gly Lys Phe 
1345                1350                1355                1360 

Gly Arg Cys Ile Asn Gln Thr Glu Gly Asp Leu Pro Leu Asn Tyr Thr 
                1365                1370                1375 

Ile Val Asn Asn Lys Ser Gln Cys Glu Ser Leu Asn Leu Thr Gly Glu 
            1380                1385                1390 

Leu Tyr Trp Thr Lys Val Lys Val Asn Phe Asp Asn Val Gly Ala Gly 
        1395                1400                1405 

Tyr Leu Ala Leu Leu Gln Val Ala Thr Phe Lys Gly Trp Met Asp Ile 
    1410                1415                1420 

Met Tyr Ala Ala Val Asp Ser Arg Gly Tyr Glu Glu Gln Pro Gln Trp 
1425                1430                1435                1440 

Glu Tyr Asn Leu Tyr Met Tyr Ile Tyr Phe Val Ile Phe Ile Ile Phe 
                1445                1450                1455 

Gly Ser Phe Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn 
            1460                1465                1470 

Phe Asn Gln Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr 
        1475                1480                1485 

Glu Glu Gln Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys 
    1490                1495                1500 

Lys Pro Gln Lys Pro Ile Pro Arg Pro Leu Asn Lys Tyr Gln Gly Phe 
1505                1510                1515                1520 

Ile Phe Asp Ile Val Thr Lys Gln Ala Phe Asp Val Thr Ile Met Phe 
                1525                1530                1535 

Leu Ile Cys Leu Asn Met Val Thr Met Met Val Glu Thr Asp Asp Gln 
            1540                1545                1550 

Ser Pro Glu Lys Ile Asn Ile Leu Ala Lys Ile Asn Leu Leu Phe Val 
        1555                1560                1565 

Ala Ile Phe Thr Gly Glu Cys Ile Val Lys Leu Ala Ala Leu Arg His 
    1570                1575                1580 

Tyr Tyr Phe Thr Asn Ser Trp Asn Ile Phe Asp Phe Val Val Val Ile 
1585                1590                1595                1600 

Leu Ser Ile Val Gly Thr Val Leu Ser Asp Ile Ile Gln Lys Tyr Phe 
                1605                1610                1615 

Phe Ser Pro Thr Leu Phe Arg Val Ile Arg Leu Ala Arg Ile Gly Arg 
            1620                1625                1630 

Ile Leu Arg Leu Ile Arg Gly Ala Lys Gly Ile Arg Thr Leu Leu Phe 
        1635                1640                1645 

Ala Leu Met Met Ser Leu Pro Ala Leu Phe Asn Ile Gly Leu Leu Leu 
    1650                1655                1660 

Phe Leu Val Met Phe Ile Tyr Ser Ile Phe Gly Met Ala Asn Phe Ala 
1665                1670                1675                1680 

Tyr Val Lys Trp Glu Ala Gly Ile Asp Asp Met Phe Asn Phe Gln Thr 
                1685                1690                1695 

Phe Ala Asn Ser Met Leu Cys Leu Phe Gln Ile Thr Thr Ser Ala Gly 
            1700                1705                1710 

Trp Asp Gly Leu Leu Ser Pro Ile Leu Asn Thr Gly Pro Pro Tyr Cys 
        1715                1720                1725 

Asp Pro Thr Leu Pro Asn Ser Asn Gly Ser Arg Gly Asp Cys Gly Ser 
    1730                1735                1740 

Pro Ala Val Gly Ile Leu Phe Phe Thr Thr Tyr Ile Ile Ile Ser Phe 
1745                1750                1755                1760 

Leu Ile Val Val Asn Met Tyr Ile Ala Ile Ile Leu Glu Asn Phe Ser 
                1765                1770                1775 

Val Ala Thr Glu Glu Ser Thr Glu Pro Leu Ser Glu Asp Asp Phe Asp 
            1780                1785                1790 

Met Phe Tyr Glu Ile Trp Glu Lys Phe Asp Pro Glu Ala Thr Gln Phe 
        1795                1800                1805 

Ile Glu Tyr Ser Val Leu Ser Asp Phe Ala Asp Ala Leu Ser Glu Pro 
    1810                1815                1820 

Leu Arg Ile Ala Lys Pro Asn Gln Ile Ser Leu Ile Asn Met Asp Leu 
1825                1830                1835                1840 

Pro Met Val Ser Gly Asp Arg Ile His Cys Met Asp Ile Leu Phe Ala 
                1845                1850                1855 

Phe Thr Lys Arg Val Leu Gly Glu Ser Gly Glu Met Asp Ala Leu Lys 
            1860                1865                1870 

Ile Gln Met Glu Glu Lys Phe Met Ala Ala Asn Pro Ser Lys Ile Ser 
        1875                1880                1885 

Tyr Glu Pro Ile Thr Thr Thr Leu Arg Arg Lys His Glu Glu Val Ser 
    1890                1895                1900 

Ala Met Val Ile Gln Arg Ala Phe Arg Arg His Leu Leu Gln Arg Ser 
1905                1910                1915                1920 

Leu Lys His Ala Ser Phe Leu Phe Arg Gln Gln Ala Gly Ser Gly Leu 
                1925                1930                1935 

Ser Glu Glu Asp Ala Pro Glu Arg Glu Gly Leu Ile Ala Tyr Val Met 
            1940                1945                1950 

Ser Glu Asn Phe Ser Arg Pro Leu Gly Pro Pro Ser Ser Ser Ser Ile 
        1955                1960                1965 

Ser Ser Thr Ser Phe Pro Pro Ser Tyr Asp Ser Val Thr Arg Ala Thr 
    1970                1975                1980 

Ser Asp Asn Leu Gln Val Arg Gly Ser Asp Tyr Ser His Ser Glu Asp 
1985                1990                1995                2000 

Leu Ala Asp Phe Pro Pro Ser Pro Asp Arg Asp Arg Glu Ser Ile Val 
                2005                2010                2015