Patent Publication Number: US-2003229062-A1

Title: Treatments for age-related macular degeneration (AMD)

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS  
     [0001] The present invention claims priority to U.S. Nonprovisional Patent Application No. 10/313,641, filed Dec. 6, 2002; U.S. Provisional Patent Application No. 60/340,498, filed Dec. 7, 2001; and U.S. Provisional Patent Application No. 60/415,864, filed Oct. 3, 2002, all of which are incorporated by reference herein in their entirety. 
    
    
     
       FIELD OF THE INVENTION  
       [0002] The present invention is directed to the fields of ophthalmology and cell biology. Specifically, the invention regards treatment of age-related macular degeneration (AMD) utilizing regulation of reverse cholesterol transport.  
       BACKGROUND OF THE INVENTION  
       [0003] Age-related macular degeneration (AMD) is the leading cause of severe visual loss in the developed world (Taylor et al., 2001; VanNewkirk et al., 2000). In the early stages of the disease, before visual loss occurs from choroidal neovascularization, there is progressive accumulation of lipids in Bruch&#39;s membrane (Pauleikhoff et al., 1990; Holz et al., 1994; Sheraidah et al., 1993; Spaide et al., 1999). Bruch&#39;s membrane lies at the critical juncture between the outer retina and its blood supply, the choriocapillaris. Lipid deposition causes reduced hydraulic conductivity and macromolecular permeability in Bruch&#39;s membrane and is thought to impair retinal metabolism (Moore et al., 1995; Pauleikhoff et al., 1990; Starita et al., 1996). Retina and/or RPE may respond by elaboration of angiogenic factors (e.g. VEGF, vFGF) that promote growth of choroidal neovascularization.  
       [0004] Interestingly, lipid accumulation in Bruch&#39;s membrane similar to that in AMD has been observed in apolipoprotein E (apo E) null mice (Dithmar et al., 2000; Kliffen et al., 2000). Because of the additional association between apo E alleles and other age-related degenerations, Alzheimer&#39;s disease and atherosclerosis, there has been recent investigation into a potential role for apo E in AMD.  
       [0005] Several studies on apo E polymorphism in AMD have been conducted (Simonelli et al., 2001; Klaver et al., 1998; Souied et al., 1998). In contrast to Alzheimer&#39;s disease, the apo E-4 allele has been associated with reduced prevalence of AMD. Apo E-2 allele is slightly increased in patients with AMD. Further supporting a role in AMD pathogenesis, apo E has been detected in drusen, the Bruch&#39;s membrane deposits that are the hallmark of AMD (Klaver et al., 1998; Anderson et al., 2001). Immunohistochemistry on post-mortem eyes has demonstrated apo E in the basal aspect of the retinal pigment epithelium (RPE) (Anderson et al., 2001). Cultured RPE cells synthesize high levels of apo E mRNA, comparable to levels found in brain (Anderson et al., 2001).  
       [0006] While the role of apo E in AMD is not established, this apolipoprotein has several functions that may affect the course of this disease. Apo E has anti-angiogenic (Browning et al., 1994), anti-inflammatory (Michael et al., 1994), and anti-oxidative effects (Tangirala et al., 2001). These are all considered atheroprotective attributes of Apo E, but may also be important in protecting against progression of AMD. While atheroprotective effects of apo E were initially thought to stem from effects on plasma lipid levels, local effects on vascular macrophages are probably equally important. Thus, selective enhanced expression of macrophage apo E in the arterial wall reduces atherosclerosis in spite of hyperlipidemia (Shimano et al., 1995; Bellosta et al., 1995; Hasty et al., 1999). Conversely, reconstitution of apo E null macrophages in C57BL/6 wild type mice induces atherosclerosis (Fazio et al., 1994). Atheroprotective effects of arterial apo E expression are thought to derive in part from facilitation of reverse cholesterol transport (Mazzone et al., 1992; Lin et al., 1999). The mechanisms by which apo E facilitates reverse cholesterol transport are incompletely understood. Apo E expression increases cholesterol efflux to HDL3 in J774 macrophages (Mazzone and Reardon, 1994) and lipid free apolipoprotein A1 (Langer et al., 2000). Cell surface apo E is also hypothesized to induce efflux from the plasma membrane (Lin et al., 1999).  
       [0007] Reverse cholesterol transport may be important in the pathogenesis of AMD because of lipid efflux from RPE into Bruch&#39;s membrane. Very much like intimal macrophages, RPE cells progressively accumulate lipid deposits throughout life; however, unlike vessel wall macrophages, the source of RPE lipid is thought to be retinal photoreceptor outer segments (POS) (Kennedy et al., 1995). Every day, each RPE cell phagocytoses and degrades more than one thousand POS via lyzosmal enzymes. These POS are enriched in phospholipid and contain the photoreactive pigment, rhodopsin. Incompletely digested POS accumulate as lipofuscin in RPE. By age 80, approximately 20% of RPE cell volume is occupied by lipofuscin (Feeney-Burns et al., 1984).  
       [0008] Analysis of Bruch&#39;s membrane lipid reveals an age-related accumulation of phospholipid, triglyceride, cholesterol, and cholesterol ester (Holz et al., 1994; Curcio et al., 2001). The origin of these lipids also is thought to derive principally from POS rather than from the circulation (Holz et al., 1994; Spaide et al., 1999). POS lipids are hypothesized to efflux from the RPE into Bruch&#39;s membrane. Although cholesterol ester deposition in Bruch&#39;s suggests contribution from plasma lipids, biochemical analysis of these esters suggests esterification of intracellular cholesterol by RPE cell derived ACAT (Curcio et al., 2002). While trafficking of lipids from the retina to RPE cells has been studied extensively, mechanisms of lipid efflux from RPE to Bruch&#39;s membrane are not well understood. Furthermore, from a pathogenic standpoint, regulation of lipid efflux into Bruch&#39;s membrane may be important in determining the rate of lipid-induced thickening that occurs in aging.  
       [0009] Nuclear hormone receptor ligands regulate reverse cholesterol transport in macrophages via their effects on ABCA-1 and apo E expression. Liver X receptor (LXR) and/or retinoid X receptor (RXR) ligands increase levels of these transporters and increase reverse cholesterol transport in macrophages (Mak et al., 2002; Laffitte et al., 2001). Thyroid hormone has also been demonstrated to increase expression of apo E three fold in HepG2 cells (Laffitte et al., 1994).  
       [0010] In AS, similar to AMD, lipids accumulate in the extracellular matrix and within phagocytic cells, primarily macrophages. Mechanisms of lipid metabolism in AS have been investigated in detail. Similar investigations into lipid processing by RPE and subsequent lipid efflux into BM and the circulation have not been conducted with the same depth as those for AS. As a consequence, potential therapeutic approaches to dry AMD are wonting.  
       [0011] Mullins et al. (2000) describe compositional similarity between drusen and other extracellular deposits, including atherosclerotic plaques. Specifically, vitronectin, amyloid P, Apo E, and lipids are among the constituents shared in common. More specifically, apolipoprotein E is identified in retinal pigmented epithelium.  
       [0012] Friedman (2000) reviews the role of atherosclerosis in the pathogenesis of AMD. Specifically, the review mentions targeting the angiogenesis pathway for treating the neovascular form of AMD, such as the member VEGF. It is noted that interfering with the upregulation or action of angiogenic agents may prove helpful for choroidal neovascularization, and, in alternative embodiments, statins may be useful for lowering the risk of AMD.  
       [0013] Anderson et al. (2001) reports apolipoprotein E protein is found in the same location as drusen, likely originating from the retinal pigmented epithelium.  
       [0014] U.S. Pat. No. 6,071,924 regards inhibition of proliferation of retinal pigment epithelium by contacting RPE cells with a retinoic acid receptor agonist, except for retinoic acid, preferably thereby inhibiting AP-1-dependent gene expression. In specific embodiments, an AP1 antagonist is delivered to a subject in need thereof for inhibition of proliferation of retinal pigment epithelium or a disease associated therewith. The related U.S. Pat. No. 6,075,032 is directed to inhibition of choroidal neovascularization by contacting RPE cells with an AP-1 antagonist. The related U.S. Pat. No. 5,824,685 regards amelioration of proliferative vitreoretinopathy or traction retinal detachment by contacting RPE cells with a retinoic acid receptor selected from ethyl-6-[2-(4,4-dimethylthiochroman-6-yl) ethynyl]nicotinate, 6-[2-(4,4-dimethylchroman-6-yl)ethynyl]nicotinic acid, and p-[(E)-2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyl)propenyl]-benzoic acid. The related U.S. Pat. No. 6,372,753 addresses inhibition of an ocular disease resulting from proliferation of retinal pigment epithelium by providing at least one AP-1 antagonist and at least one retinoic acid receptor (RAR) agonist, except for retinoic acid.  
       [0015] WO 01/58494 is directed to treating or preventing an ocular disease, such as age-related macular degeneration, by contacting an ocular cell with an expression vector comprising a nucleic acid sequence encoding an inhibitor of angiogenesis and a neurotrophic agent. In specific embodiments, the inhibitor of angiogenesis and the neurotrophic agent are one and the same, such as pigment epithelium-derived factor (PEDF).  
       [0016] WO 02/13812 regards the use of an insulin-sensitizing agent, preferably peroxisome proliferator-activated receptor-γ (PPAR γ) agonists, for the treatment of an inflammatory disease, such as an ophthalmic disease.  
       [0017] WO 00/52479 addresses diagnosing, treating, and preventing drusen-associated disorders (any disorder which involves drusen formation), including AMD. In specific embodiments, there are methods related to providing an effective amount of an agent that inhibits immune cell proliferation or differentiation, such as antagonists of TNF-alpha.  
       [0018] Thus, the present invention provides a novel approach to reduce lipid content of ocular tissue, such as Bruch&#39;s membrane and further provides methods and compositions for the treatment of macular degeneration, such as AMD.  
       BRIEF SUMMARY OF THE INVENTION  
       [0019] The present invention is directed to a system, method, and/or composition(s) related to treating AMD. Treatments for dry AMD have been lacking, because the pathogenesis of this common condition is poorly understood, and the inventors have demonstrated analogous biological behavior between human retinal pigment epithelial (RPE) cells and macrophages that point toward similar pathogenic mechanisms of AMD and atherosclerosis. Specifically, reverse cholesterol transport (RCT) is exploited in the present invention for the treatment of AMD. The present inventors provide the novel demonstration of RCT in RPE cells in the eye. More specifically, RCT is regulated through manipulation of levels of cholesterol and/or phospholipid transporters (ABCA-1, Apo E, SRB-1, SRB-2) by nuclear hormone receptor ligands such as agonists of thyroid hormone (TR), liver X receptor (LXR), and/or retinoid X receptor (RXR). A goal for the present invention is the reduction of lipid content of RPE Bruch&#39;s membrane to facilitate an improvement in visual function and/or, in some embodiments, prevent ocular disease, such as AMD. Reduction of the lipid content of Bruch&#39;s membrane preferably results in at least one or more of the following: reduction in development of CNV; improvement in dark adaptation; improvement in night vision; improved visual acuity; and/or improved recovery to bright flash stimulus.  
       [0020] In a specific embodiment of the present invention, patients with drusen and no evidence of choroidal neovascularization are administered a nuclear hormone agonist, such as thyroid hormone (TR) agonist (for example, T3 (3,5,3′-L-triiodothyronine), TRIAC (3-triiodothyoacetic acid), GC1, KB-000,141 and/or KB141 (Karo Bio; Huddinge, Sweden). Administration could be orally or by sustained release systems well known in the art. In a specific embodiment, the agonist binds to at least one nuclear hormone receptor in the RPE and induces upregulation of RCT. Efflux of lipid from RPE increases, and the likelihood of visual loss from choroidal neovascularization is reduced. Other nuclear hormone receptor ligands of the TR, RXR, and LXR families, for example, which are well known in the art, are used independently or in combination with each other to enhance RCT by RPE.  
       [0021] In an embodiment of the present invention, there is a method of increasing lipid efflux from an ocular tissue, comprising the step of delivering to the tissue a nuclear hormone receptor ligand. In a specific embodiment, ocular tissue is retinal pigment epithelium (RPE), Bruch&#39;s membrane, or a combination thereof. In another specific embodiment, the nuclear hormone receptor is thyroid hormone receptor. In an additional specific embodiment, the ligand of thyroid hormone receptor is 3,5,3′-L-triiodothyronine (T3), TRIAC (3-triiodothyroacetic acid); KB141; GC-1; 3, 5 dimethyl-3-isopropylthyronine; or a mixture thereof. In one embodiment, the nuclear hormone receptor is liver X receptor. In a specific embodiment, the ligand of liver X receptor is 22 (R) hydroxycholesterol; acetyl-podocarpic dimer; T0901317; GW3965 (12); 24(S),25-epoxycholesterol; 24(R),25-epoxycholesterol; 22(R)-ol-24(S),25-epoxycholesterol; 22(S)-ol,24(R),25-epoxycholesterol; 24(S),25-iminocholesterol; methyl-H-cholenate; dimethyl-hydroxycholenamide; 24(S)-hydroxycholesterol; 24(R)-hydroxycholesterol; 22(S)-hydroxycholesterol; 22(R),24(S)-dihydroxycholesterol; 25-hydroxycholesterol; 24(S),25-dihydroxycholesterol; 24(R),25-dihydroxycholesterol; 24,25-dehydrocholesterol; 7(a)-ol, 24(S),25-epoxycholesterol; 7(b)-ol,24(S),25-epoxycholesterol; 7k,24(S),25-epoxycholesterol; 7(α)-hydroxycholesterol; 7-ketocholesterol; cholesterol; 5,6-24(S),25-diepoxycholesterol; or a mixture thereof. In a specific embodiment, the nuclear hormone receptor is retinoid X receptor, ligands of which include 9 cis-retinoic acid; AGN 191659 [(E)-5-[2-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-naphthyl)propen-1-yl]-2-thiophenecarboxylic acid]; AGN 191701 [(E) 2-[2-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-naphthyl)propen-1-yl]-4-thiophene-carboxylic acid]; AGN 192849 [(3,5,5,8,8,-pentamethyl-5,6,7,8-tetrahydronaphthalen-2-yl) (5 carboxypyrid-2-yl)sulfide]; LGD346; LG100268; LG100754; BMS649; bexaroteneR (4-[1-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-naphthalenyl) ethenyl] benzoic acid); or a mixture thereof. In one embodiment, the ocular tissue is comprised in an individual, such as one at risk for developing macular degeneration or another ocular disease, and/or the ocular tissue is comprised in individuals afflicted with macular degeneration (for example, age-related macular degeneration). In other specific embodiment, the individual is afflicted with Stargardts disease ( fundus flavimaculatus ) or is at risk for developing Stargardts disease.  
       [0022] In another embodiment of the present invention, there is a method of increasing reverse cholesterol transport in an ocular tissue, comprising the step of delivering to the tissue at least one ligand of a nuclear hormone receptor. In a specific embodiment, the ocular tissue is retinal pigment epithelium (RPE), Bruch&#39;s membrane, or a combination thereof.  
       [0023] In an additional embodiment of the present invention, there is a method of treating macular degeneration (AMD) in an individual, comprising the step of delivering to the individual a ligand of a nuclear hormone receptor. In a specific embodiment, the delivering occurs under conditions wherein reverse cholesterol transport is upregulated.  
       [0024] In another embodiment of the present invention, there is a kit for the treatment of macular degeneration, housed in a suitable container, comprising a ligand of a nuclear hormone receptor. In a specific embodiment, the nuclear hormone receptor is TR, RXR, LXR, or a combination thereof. In some embodiments, it is useful to comprise a combination of nuclear hormone receptors, since it is well known in the art that many are heterodimers (for example, LXR and RXR, or RXR and TR). In a specific embodiment, the kit comprises a pharmaceutically acceptable excipient. In another specific embodiment, the ligand for a nuclear hormone receptor is comprised in the pharmaceutically acceptable excipient.  
       [0025] In an additional embodiment of the present invention, there is a method of treating macular degeneration (AMD) in an individual, comprising the steps of identifying a ligand for a nuclear hormone receptor; and delivering said ligand to said individual. Methods to identify a ligand for a nuclear hormone receptor are well known in the art.  
       [0026] The foregoing has outlined rather broadly the features and technical advantages of the present invention in order that the detailed description of the invention that follows may be better understood. Additional features and advantages of the invention will be described hereinafter which form the subject of the claims of the invention. It should be appreciated by those skilled in the art that the conception and specific embodiment disclosed may be readily utilized as a basis for modifying or designing other structures for carrying out the same purposes of the present invention. It should also be realized by those skilled in the art that such equivalent constructions do not depart from the spirit and scope of the invention as set forth in the appended claims. The novel features which are believed to be characteristic of the invention, both as to its organization and method of operation, together with further objects and advantages will be better understood from the following description when considered in connection with the accompanying figures. It is to be expressly understood, however, that each of the figures is provided for the purpose of illustration and description only and is not intended as a definition of the limits of the present invention. 
     
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
     [0027] For a more complete understanding of the present invention, reference is now made to the following descriptions taken in conjunction with the accompanying drawing, in which:  
     [0028]FIG. 1 shows that RPE cells express Apo E, ABCA1, and LXR α.  
     [0029]FIG. 2 shows RPE cell expression of SR-BI and SR-BII.  
     [0030]FIG. 3 illustrates SR-BI and SR-BII immunofluorescence in RPE cells.  
     [0031]FIG. 4 demonstrates ABCA1 immunofluorescence in RPE cells.  
     [0032]FIG. 5 demonstrates that basal Apo E expression is greater than apical Apo E expression in cultured human RPE cells.  
     [0033]FIG. 6 shows regulation of Apo E expression by nuclear hormone receptor ligands.  
     [0034]FIG. 7 provides a non-denatured polyacrylamide gel of lipoprotein fractions.  
     [0035]FIG. 8 shows  14 C distributeion of the fractions from FIG. 7.  
     [0036]FIG. 9 demonstrates thin layer chromatography illustrating the identification of six out of seventeen spots of an HDL fraction. Note: HDL is the high density lipoprotein fraction; POS is labeled POS starting material; PC is phophatidylcholine; PI is phosphatidylinisotol; PE is phosphatidylethanolamine; C is cholesterol; TRL is TG rich lipid, including triglycerides and cholesterol ester.  
     [0037]FIG. 10 demonstrates that  14 C counts increase following drug treatments that increase RCT.  
     [0038]FIG. 11 illustrates ABCA1 regulation by RXR and LXR ligands.  
     [0039]FIG. 12 shows HDL, LDL and plasma stimulation of  14 C-labeled lipid transport the identification of HDL from RPE cells.  
     [0040]FIG. 13 shows stimulation of CD36 expression by oxidized lipid.  
     [0041]FIG. 14 illustrates apical and basal secretion from RPE cells of apoE in the presence of T 3  (T), 22(R) hydroxycholesterol, or cis retinoic acid (RA).  
     [0042]FIG. 15 shows that apoE secreted from RPE cells binds to HDL.  
     [0043]FIG. 16 demonstrates that HDL stimulates lipid efflux from RPE cells in culture.  
     [0044]FIG. 17 shows characterization of HDL and plasma bound POS lipids by thin layer chromatography.  
     [0045]FIG. 18 shows plasma and HDL levels of species identified in FIG. 17. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION  
     [0046] I. Definitions  
     [0047] As used herein the specification, “a” or “an” may mean one or more. As used herein in the claim(s), when used in conjunction with the word “comprising”, the words “a” or “an” may mean one or more than one. As used herein “another” may mean at least a second or more.  
     [0048] The term “age-related macular degeneration” as used herein refers to macular degeneration in an individual over the age of about 50. In one specific embodiment, it is associated with destruction and loss of the photoreceptors in the macula region of the retina resulting in decreased central vision and, in advanced cases, legal blindness.  
     [0049] The term “Bruch&#39;s membrane” as used herein refers to a five-layered structure separating the choriocapillaris from the RPE.  
     [0050] The term “HDL or subspecies thereof” refers to the fact that high density lipoproteins (HDL) can be fractionated into particulate species defined in molecular size and composition. HDL as prepared by density ultracentrifugation and by native nondenaturing purification processes including anti-apolipoprotein A-I immunoaffinity chromatography have been characterized for its constituent species by two-dimensional nondenaturing polyacrylamide electrophoresis, immunoblotting, and mass spectroscopy. HDL has been resolved into more than twenty-five particle species that differ in charge and molecular size. Each particle is defined by a unique combination of protein (including apolipoproteins A-I, A-II, A-IV, A-V, C-III, D, E, J, L, lecithin:cholesterol acyltransferase, cholesterol ester transferase, phospholipid transfer protein, alpha-2 macroglobulin) and lipid (including phospholipid, triglyceride, cholesterol, cholesterol ester, fatty acids). A partial list of HDL species include HDL alpha-1, HDL alpha-2, HDL alpha-3, HDL prebeta-1, HDL prebeta-2 (and variants “a”, “b”, “c”, “d”), HDL prebeta-3, HDL prebeta-4, and HDL prealpha-1.  
     [0051] The term “increase lipid efflux” or “increasing lipid efflux” as used herein refers to an increased level and/or rate of lipid efflux, promoting lipid efflux, enhancing lipid efflux, facilitating lipid efflux, upregulating lipid efflux, improving lipid efflux, and/or augmenting lipid efflux. In a specific embodiment, the efflux comprises efflux of phospholipid, triglyceride, cholesterol, and/or cholesterol ester.  
     [0052] A skilled artisan recognizes that the term “lipid transporter” as used herein refers to a lipoprotein that carries lipids away from peripheral cells into the circulation, and examples include HDL and subspecies thereof, or a mixture thereof. The term “lipid transporter” is also used in the art to refer to, for example, transmembrane proteins that transport cholesterol and phospholipids, for example, from inside a cell to outside the cell. Examples include ABCA1, SR-BI, SR-BII, ABCA4, ABCG5, ABCG8, or a mixture thereof.  
     [0053] The term “macula” as used herein refers to the light-sensing cells of the central region of the retina.  
     [0054] The term “macular degeneration” as used herein refers to deterioration of the central portion of the retina, the macula.  
     [0055] The term “nuclear hormone receptor” as used herein refers to an intracellular receptor that plays a role in expression of gene(s) involved in physiological processes, examples of which include cell growth and differentiation, development, and homeostasis. In a specific embodiment, these receptors are members of a superfamily of receptors, whose members recognize similar DNA sequences that contain two or more hexanucleotide DNA-binding half-sites arranged as direct repeats or inverted repeats. It is through this recognition that these receptors are able to regulate the expression of genes in the nucleus, and thereby regulate the respective physiological process. Examples of nuclear hormone receptors include thyroid hormone receptor, liver X receptor, retinoid X receptor, estrogen receptor, androgen receptor, peroxisome proliferator activated receptors (PPARs), trans-retinoic acid receptor (RAR), the vitamin D receptor (VDR), glucocorticoid receptor, the progesterone receptor, and isoforms thereof.  
     [0056] Nuclear hormone receptors in some embodiments include those that remain sequestered in the cytoplasm in the absence of their cognate ligands (e.g., steroid hormone receptors). Upon binding of the ligand, the steroid hormone receptors are translocated to the nucleus where they bind to hormone response elements, typically as homodimers.  
     [0057] In other embodiments the nuclear hormone receptors are not sequestered in the cytoplasm in the absence of their ligands, but rather remain in the nucleus. These receptors, which include the thyroid hormone, retinoid, fatty acid, and eicosanoid receptors, typically bind to their cognate response elements as heterodimers with, for example, a 9-cis-retinoic acid receptor (RXR). Often, binding of a nuclear receptor to a response element occurs in the absence of the cognate ligand. An example of such a nuclear receptor is the farnesoid X receptor (FXR).  
     [0058] Methods to identify ligands of nuclear hormone receptors are well known in the art, examples of which are described in U.S. Pat. No. 5,846,711 and U.S. Pat. No. 6,266,622, both of which are incorporated by reference herein in their entirety.  
     [0059] The term “reverse cholesterol transport” as used herein refers to transport of cholesterol from peripheral tissues to the liver. In a specific embodiment, it refers to efflux of lipid from RPE cells. In specific embodiments, it comprises efflux of cellular cholesterol and/or phospholipid to HDL, and, in further specific embodiments, it comprises HDL delivery of cholesterol ester to the liver, such as for biliary secretion.  
     [0060] The term “therapeutically effective” as used herein refers to the amount of a compound required to improve some symptom associated with a disease. For example, in the treatment of macular degeneration, a compound which improves sight to any degree or arrests any symptom of impaired sight would be therapeutically effective. A therapeutically effective amount of a compound is not required to cure a disease but will provide a treatment for a disease.  
     [0061] The term “upregulate” as used herein is defined as increasing the level and/or rate of an event, process, or mechanism, such as reverse cholesterol transport and/or the transcription and or translation processes of a nucleic acid, such as a gene.  
     [0062] II. The Present Invention  
     [0063] As stated, the histopathology of macula in patients with AMD shows diffuse thickening of Bruch&#39;s membrane, and the overlying RPE is attenuated and full of lipofuscin granules. Photoreceptors are shortened and atrophic, and much of the thickened Bruch&#39;s membrane consists of lipid deposition. It is known that following about 50 years of age, the rate of lipid accumulation accelerates (Holz et al., 1994).  
     [0064] Using cell culture methods to study lipid metabolism, the inventors have shown a number of analogous mechanisms for lipid metabolism that are shared by macrophages and human RPE cells. The shared biology of these two cell types indicates useful therapeutic approaches for treatment of AMD. Specifically, the present inventors are the first to show that RCT occurs in RPE cells, and enhancement of RCT is beneficial for removing undesired lipid from the RPE cells and/or Bruch&#39;s membrane to facilitate retinal metabolism. In a specific embodiment, the transporters in the RCT system are regulated to improve RCT. In a further specific embodiment, this regulatory aspect of the present invention provides a novel treatment for AMD.  
     [0065] Although there has been discussion in the field regarding mechanisms of lipid accumulation in macula of AMD individuals, the present invention regards efflux of lipid into the circulation, which reduces the amount of lipid in RPE and/or Bruch&#39;s membrane. Promotion of this efflux comprises one aspect of the invention and is an effective therapy for both early and late AMD. A skilled artisan recgonizes that early AMD comprises the presence of drusen and late stage AMD comprises visual loss from choroidal neovascularization or geographic atrophy.  
     [0066] Thus, the present invention provides the novel idea in the field in which reverse cholesterol transport occurs in RPE cells. In specific embodiments, the invention provides methods and compositions related to facilitating efflux of cholesterol and/or phospholipids from inside an RPE cell to the outside of the RPE cell, and further through Bruch&#39;s membrane. In another specific embodiment, following efflux from Bruch&#39;s membrane the cholesterol and/or phospholipids are transported by apolipoprotein E, apolipoprotein A1, and other transporters, or a combination thereof, to HDL for removal to the liver.  
     [0067] A skilled artisan recognizes the important role reverse cholesterol transport (RCT) plays in lipid homeostasis. HDL levels are inversely correlated with incidence of coronary artery disease (CAD). Tangier&#39;s disease, which comprises a mutation of ABCA1, leads to deposition of cholesterol in reticuloendothelial tissues and premature atherosclerosis. Furthermore, the Apo E null mouse is an excellent model of atherosclerosis and hyperlipidemia. Interestingly, supporting an important role of Apo E in RCT, reconstitution of Apo E positive macrophages via bone marrow transplant into an Apo E null mouse prevents atherosclerosis. This occurs in spite of persistent hyperlipidemia.  
     [0068] In one embodiment of the present invention a transporter of lipid from RPE cells is enhanced for the transport activity, such as by an increase in the level of the transporter. Examples of transporters include apo E, ABCA1, SR-BI, SR-BII, ABCA4, ABCG5, ABCG8; other proteins that might be involved are LCAT, CETP, PLTP, LRP receptor, LDL receptor, Lox-1, and lipases. In a specific embodiment, lox-1 and PLTP are expressed in RPE, as demonstrated by RT_PCR. In a specific embodiment of the present invention, apo A1 is utilized to facilitate RCT from RPE cells. In an additional specific embodiment, apo A1 is made by RPE cells.  
     [0069] In a specific embodiment of the present invention, strategies for intervention for treatment of AMD are provided in which reverse cholesterol transport is enhanced at the level of the RPE by upregulating ABCA1, Apo E, SR-BI and/or SR-BII expression. SR-B has been reported to be upregulated by 17beta-Estradiol and testosterone. Additionally, or alone, HDL binding to effluxed lipids is enhanced, thereby increasing efflux of lipids from Bruch&#39;s membrane into the circulation and providing therapy for AMD. In one embodiment, an increase in HDL levels is utilized to facilitate lipid efflux from RPE cells and/or Bruch&#39;s membrane, and in a specific embodiment, levels of specific subspecies of HDL are utilized to facilitate lipid efflux. For example, effluxed lipids could bind to preβ-HDL, HDL1, HDL2 or HDL3. Effluxed lipids could also bind prebeta-1, prebeta-2, prebeta-3, and/or prebeta-4 HDL. In a specific embodiment, the effluxed lipids bind preferentially to HDL2 that comprises apo E.  
     [0070] One skilled in the art recognizes particular RCT components are present in RPE cells (Mullins et al., 2000; Anderson et al., 2001). Nuclear hormone receptors known to regulate expression of reverse cholesterol transport proteins are also expressed in cultured human RPE. Thus, in a preferred embodiment of the present invention, ligands to at least one of the nuclear hormone receptors upregulates RCT. In further embodiments, following efflux from RPE cells, the lipids bind HDL, so in an embodiment of the present invention there is upregulation of HDL for AMD treatment, such as by statins and/or niacin.  
     [0071] In an alternative embodiment, treatment for AMD comprises reduction of RCT. For example, in individuals past a certain age, such as about 50, 55, 60, 65, 70, 75, 80, and so on, the transporters are preferentially inhibited. In one aspect of this embodiment, HDL is unable to enter Bruch&#39;s membrane to remove the lipids and the RPE continues to efflux lipids. In such cases where effluxed lipids from RPE cannot be removed by a lipoprotein acceptor, lipid efflux by RPE is inhibited to maintain macromolecular transport across Bruch&#39;s membrane. Inhibition of RCT by reducing levels of ABCA-1, apo E, and/or SRB-1, or SRB-2 would reduce accumulation of lipid in Bruch&#39;s membrane.  
     [0072] In embodiments of the present invention, ligands for nuclear hormone receptors are utilized as compounds for enhancing RCT for the reduction of lipid content of RPE and Bruch&#39;s membrane. In a specific embodiment, the nuclear hormone receptor ligands are utilized for treatment of AMD. In a further specific embodiment, the nuclear hormone receptors comprise TR, RXR, and/or LXR. In other specific embodiments, ligands of the nuclear hormone receptors are delivered to at least one RPE cell to facilitate efflux of lipids from the RPE cell and/or are delivered to Bruch&#39;s membrane for efflux from Bruch&#39;s membrane. Examples of ligands for TR include T3 (3,5,3′-L-triiodothyronine). Other examples of TR ligands include but are not limited to TRIAC (3-triiodothyroacetic acid); KB141 (Karo Bio); GC-1; and 3, 5 dimethyl-3-isopropylthyronine. Examples of ligands for RXR include 9 cis-retinoic acid, and other RXR ligands also include but are not limited to: AGN 191659 [(E)-5-[2-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-naphthyl)propen-1-yl]-2-thiophenecarboxylic acid]; AGN 191701 [(E) 2-[2-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-naphthyl)propen-1 -yl]-4-thiophene-carboxylic acid]; AGN 192849 [(3,5,5,8,8,-pentamethyl-5,6,7,8-tetrahydronaphthalen-2-yl) (5 carboxypyrid-2-yl)sulfide]; LGD346; LG100268; LG100754; BMS649; and bexaroteneR (Ligand Pharmaceuticals) (4-[1-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-naphthalenyl) ethenyl] benzoic acid). Examples of ligands for LXR include 22 (R) hydroxycholesterol, acetyl-podocarpic dimer, T0901317, and GW3965.  
     [0073] In an embodiment of the present invention, expression of a sequence is monitored following administration of an upregulator of its expression or a compound suspected to be an upregulator. A skilled artisan recognizes how to obtain these sequences, such as commercially from Celera Genomics, Inc. (Rockville, Md.) or from the National Center for Biotechnology Information&#39;s GenBank database. Exemplary apo E polynucleotide sequences include the following, cited with their GenBank Accession number: SEQ ID NO: 1 (K00396); SEQ ID NO:2 (M10065); and SEQ ID NO:3 (M12529). Some exemplary apo E polypeptide sequences include the following, cited with their GenBank Accession number: SEQ ID NO:4 (AAB59546); SEQ ID NO:5 (AAB59397); and SEQ ID NO:6 (AAB59518).  
     [0074] In other embodiments, sequences of ABCA-1 are utilized, such as to monitor ABCA-1 expression related to methods of the present invention. Some examples of ABCA1 polynucleotides include SEQ ID NO:7 (NM — 005502); and SEQ ID NO:8 (AB055982). Some examples of ABCA1 polypeptides include SEQ ID NO:9 (NP — 005493); and SEQ ID NO:10 (BAB63210).  
     [0075] In some methods of the present invention, expression levels of sequences of SR-BI and SR-B2 polynucleotides are monitored following administration of a nuclear hormone receptor ligand. An example of SR-BI polynucleotide is SEQ ID NO:11 (NM — 005505) and an example of a SR-BI polypeptide is SEQ ID NO:12 (NP — 005496).  
     [0076] III. Pharmaceutical Compositions and Routes of Administration  
     [0077] Compositions of the present invention may have an effective amount of a compound for therapeutic administration and, in some embodiments, in combination with an effective amount of a second compound that is also an anti-AMD agent. In a specific embodiment, the compound is a ligand/agonist of a nuclear hormone receptor. In other embodiments, compounds that upregulate expression of HDL are the compounds for therapeutic administration. Such compositions will generally be dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium.  
     [0078] The phrases “pharmaceutically or pharmacologically acceptable” refer to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to an animal, or human, as appropriate. As used herein, “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredients, its use in the therapeutic compositions is contemplated. Supplementary active ingredients, such as other anti-AMD agents, can also be incorporated into the compositions.  
     [0079] In addition to the compounds formulated for parenteral administration, such as intravenous or intramuscular injection, other pharmaceutically acceptable forms include, e.g., tablets or other solids for oral administration; time release capsules; and any other form currently used, including cremes, lotions, mouthwashes, inhalants and the like.  
     [0080] The delivery vehicles of the present invention may include classic pharmaceutical preparations. Administration of these compositions according to the present invention will be via any common route so long as the target ocular tissue is available via that route. This includes oral, nasal, buccal, rectal, vaginal or topical. Alternatively, administration may be by orthotopic, intradermal, subcutaneous, intramuscular, intraperitoneal or intravenous injection. Such compositions would normally be administered as pharmaceutically acceptable compositions. In some embodiments, the compositions are administered by sustained release intra- or extra-ocular devices.  
     [0081] The vehicles and therapeutic compounds therein of the present invention are advantageously administered in the form of injectable compositions either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid prior to injection also may be prepared. These preparations also may be emulsified. A typical composition for such purposes comprises a 50 mg or up to about 100 mg of human serum albumin per milliliter of phosphate buffered saline. Other pharmaceutically acceptable carriers include aqueous solutions, non-toxic excipients, including salts, preservatives, buffers and the like. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oil and injectable organic esters, such as theyloleate. Aqueous carriers include water, alcoholic/aqueous solutions, saline solutions, parenteral vehicles such as sodium chloride, Ringer&#39;s dextrose, etc. Intravenous vehicles include fluid and nutrient replenishers. Preservatives include antimicrobial agents, anti-oxidants, chelating agents and inert gases. The pH and exact concentration of the various components in the pharmaceutical are adjusted according to well-known parameters.  
     [0082] Additional formulations are suitable for oral administration. Oral formulations include such typical excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate and the like. The compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders. When the route is topical, the form may be a cream, ointment, salve or spray.  
     [0083] An effective amount of the therapeutic agent is determined based on the intended goal. The term “unit dose” refers to a physically discrete unit suitable for use in a subject, each unit containing a predetermined quantity of the therapeutic composition calculated to produce the desired response in association with its administration, i.e., the appropriate route and treatment regimen. The quantity to be administered, both according to number of treatments and unit dose, depends on the subject to be treated, the state of the subject and the protection desired. Precise amounts of the therapeutic composition also depend on the judgment of the practitioner and are peculiar to each individual.  
     [0084] All of the essential materials and reagents required for AMD treatment, diagnosis and/or prevention may be assembled together in a kit. When the components of the kit are provided in one or more liquid solutions, the liquid solution preferably is an aqueous solution, with a sterile aqueous solution being particularly preferred.  
     [0085] For in vivo use, an anti-AMD agent may be formulated into a single or separate pharmaceutically acceptable syringeable composition. In this case, the container means may itself be an inhalant, syringe, pipette, eye dropper, or other such like apparatus, from which the formulation may be applied to an infected area of the body, such as the lungs, injected into an animal, or even applied to and mixed with the other components of the kit.  
     [0086] The components of the kit may also be provided in dried or lyophilized forms. When reagents or components are provided as a dried form, reconstitution generally is by the addition of a suitable solvent. It is envisioned that the solvent also may be provided in another container means. The kits of the invention may also include an instruction sheet defining administration of the anti-AMD composition.  
     [0087] The kits of the present invention also will typically include a means for containing the vials in close confinement for commercial sale such as, e.g., injection or blow-molded plastic containers into which the desired vials are retained. Irrespective of the number or type of containers, the kits of the invention also may comprise, or be packaged with, an instrument for assisting with the injection/administration or placement of the ultimate complex composition within the body of an animal. Such an instrument may be an inhalant, syringe, pipette, forceps, measured spoon, eye dropper or any such medically approved delivery vehicle.  
     [0088] The active compounds of the present invention will often be formulated for parenteral administration, e.g. formulated for injection via the intravenous, intramuscular, subcutaneous, or even intraperitoneal routes. The preparation of an aqueous composition that contains a second agent(s) as active ingredients will be known to those of skill in the art in light of the present disclosure. Typically, such compositions can be prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for using to prepare solutions or suspensions upon the addition of a liquid prior to injection can also be prepared; and the preparations can also be emulsified.  
     [0089] Solutions of the active compounds as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.  
     [0090] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.  
     [0091] The active compounds may be formulated into a composition in a neutral or salt form. Pharmaceutically acceptable salts, include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like.  
     [0092] The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and/or antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.  
     [0093] Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.  
     [0094] In certain cases, the therapeutic formulations of the invention could also be prepared in forms suitable for topical administration, such as in eye drops, cremes and lotions.  
     [0095] Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, with even drug release capsules and the like being employable.  
     [0096] For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intraocular, intravenous, intramuscular, and subcutaneous administration. In this connection, sterile aqueous media that can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage could be dissolved in 1 mL of isotonic NaCl solution and either added to 1000 mL of hypodermoclysis fluid or injected at the proposed site of infusion, (see for example, “Remington&#39;s Pharmaceutical Sciences” 15th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.  
     [0097] Targeting of ocular tissues may be accomplished in any one of a variety of ways. In one embodiment, there is the use of liposomes to target a compound of the present invention to the eye, and preferably to RPE cells and/or Bruch&#39;s membrane. For example, the compound may be complexed with liposomes in the manner described above, and this compound/liposome complex injected into patients with AMD, using intravenous injection to direct the compound to the desired ocular tissue or cell. Directly injecting the liposome complex into the proximity of the RPE or Bruch&#39;s membrane can also provide for targeting of the complex with some forms of AMD. In a specific embodiment, the compound is administered via intra-ocular sustained delivery (such as Vitrasert® or Envision® by Bauch and). In a specific embodiment, the compound is delivered by posterior subtenons injection. In another specific embodiment, microemulsion particles with apo E (such as, recombinant) are delivered to ocular tissue to take up lipid from Bruch&#39;s membrane, RPE cells, or both.  
     [0098] Those of skill in the art will recognize that the best treatment regimens for using compounds of the present invention to treat AMD can be straightforwardly determined. This is not a question of experimentation, but rather one of optimization, which is routinely conducted in the medical arts. In vivo studies in nude mice often provide a starting point from which to begin to optimize the dosage and delivery regimes. The frequency of injection will initially be once a week, as has been done in some mice studies. However, this frequency might be optimally adjusted from one day to every two weeks to monthly, depending upon the results obtained from the initial clinical trials and the needs of a particular patient. Human dosage amounts can initially be determined by extrapolating from the amount of compound used in mice, as a skilled artisan recognizes it is routine in the art to modify the dosage for humans compared to animal models. In certain embodiments it is envisioned that the dosage may vary from between about 1 mg compound/Kg body weight to about 5000 mg compound/Kg body weight; or from about 5 mg/Kg body weight to about 4000 mg/Kg body weight or from about 10 mg/Kg body weight to about 3000 mg/Kg body weight; or from about 50 mg/Kg body weight to about 2000 mg/Kg body weight; or from about 100 mg/Kg body weight to about 1000 mg/Kg body weight; or from about 150 mg/Kg body weight to about 500 mg/Kg body weight. In other embodiments this dose may be about 1, 5, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1600, 1700, 1800, 1900, 2000, 2500, 3000, 3500, 4000, 4500, 5000 mg/Kg body weight. In other embodiments, it is envisaged that higher does may be used, such doses may be in the range of about 5 mg compound/Kg body to about 20 mg compound/Kg body. In other embodiments the doses may be about 8, 10, 12, 14, 16 or 18 mg/Kg body weight. Of course, this dosage amount may be adjusted upward or downward, as is routinely done in such treatment protocols, depending on the results of the initial clinical trials and the needs of a particular patient.  
     [0099] IV. Kits  
     [0100] Any of the compositions described herein may be comprised in a kit. In a non-limiting example, a nuclear hormone receptor agonist or ligand, and in some embodiments, at least one additional agent, may be comprised in a kit. In other embodiments, a lipid transporter such as HDL or a subspecies thereof.  
     [0101] The kits may comprise a suitably aliquoted nuclear hormone receptor ligand, and/or additional agent compositions of the present invention, whether labeled or unlabeled, as may be used to prepare a standard curve for treatment of macular degeneration, such as AMD. The components of the kits may be packaged in aqueous media or in lyophilized form. When reagents and/or components are provided as a dry powder, the powder can be reconstituted by the addition of a suitable solvent. It is envisioned that the solvent may also be provided in another container means.  
     [0102] The container means of the kits will generally include at least one vial, test tube, flask, bottle, syringe or other container means, into which a component may be placed, and preferably, suitably aliquoted. Where there are more than one component in the kit, the kit also will generally contain a second, third or other additional container into which the additional components may be separately placed. However, various combinations of components may be comprised in a vial. The kits of the present invention also will typically include a means for containing the nuclear hormone receptor ligand, additional agent, and any other reagent containers in close confinement for commercial sale. Such containers may include injection or blow-molded plastic containers into which the desired vials are retained.  
     EXAMPLES  
     [0103] The following is an illustration of preferred embodiments for practicing the present invention. However, they are not limiting examples. Other examples and methods are possible in practicing the present invention.  
     Example 1  
     MATERIALS AND METHODS  
     [0104] Cell Culture and Drug Treatments  
     [0105] Primary cultures of normal human RPE cells from passages  5  to  10  were used for the experiments described. RPE cells were grown to confluence on laminin-coated  6  well Transwell tissue culture plates (Costar) with DMEM-H21 containing 10% fetal bovine serum, 2 mM glutamine, 50 μg/ml gentamicin and 2.5 mg/ml fungizone in the top and bottom chambers. For immunofluorescent staining cells were grown on laminin coated slides in the same medium. Cells were grown for at least 1 week at confluence prior to drug treatment. Cells to be treated with drugs were incubated in serum free DMEM-H21 prior to drug addition. Drug treatments were in serum free DMEM-H21 with or without 10 −7  M thyroid hormone (T 3 ), 2.5×10 −6  M 22 (R) hydroxycholesterol, or 10 −7  M cis retinoic acid in both chambers for 36 hours.  
     [0106] RT-PCR  
     [0107] Confluent cell cultures were harvested and total RNA was purified using RNAzol (Teltest, Inc., Friendswood, Tex.) according to the manufacturer&#39;s instructions. Equal amounts of purified RNA were used in each reaction as templates for cDNA synthesis using the 1st Strand Synthesis Kit for RT-PCR (AMV) (Boehringer, Indianapolis, Ind.). RT-PCR was carried out on 1 μg of cDNA with Amplitaq Taq polymerase (Perkin-Elmer, Branchburg, N.J.). In some experiments apo E RT-PCR products were quantified using the Quantu mRNA  assay kit according to the manufacturer&#39;s instructions (Ambion, Austin, Tex.). Briefly, 18S rRNA and apo E cDNAs are simultaneously amplified in each reaction. The RT-PCR products are resolved by electrophoresis on 1.4% agarose gels. The apo E mRNA expression is assessed relative to the internal 18S rRNA expression by densitometric analysis of photographed agarose gels.  
     [0108] RT-PCR primers specific to human apo E, ABCA1, SR-BI, SR-BII, and lxr α were used. The RT-PCR product of the predicted sizes for the apo E, ABCA1, SR-BI, and SR-BII RT-PCR products were excised form the gel and their identities were confirmed by DNA sequencing (not shown).  
     [0109] Immunofluoresence Microscopy  
     [0110] RPE cells, grown on slides, were σταινεδ with either antisera to ABCA1, or with purified antibodies to SR-BI or SR-BII. Cells were fixed in ice cold 100% MeOH for 20 min. All subsequent steps were performed at room temperature. Cells were washed in phosphate buffered saline (PBS) and incubated for in 5% goat serum in PBS for 30 min. Cells were then washed in buffer A (150 mM NaCl, 10 mM phosphate, pH 7.8) and incubated with the primary antibody in buffer A for 45 min. After washing with buffer A the cells were incubated in Avidin Blocking Reagent (Vector Laboratories, Burlingame, Calif.) for 15 min, washed in buffer A again and incubated in Biotin Blocking Reagent (Vector Laboratories, Burlingame, Calif.) for 15 min. After washing in buffer A, cells were incubated in 10 μg/ml biotinylated goat anti-rabbit IgG (Vector Laboratories, Burlingame, Calif.) in buffer A for 30 min, washed in buffer A and incubated in 20 μg/ml fluorescein conjugated avidin D (Vector Laboratories, Burlingame, Calif.) in buffer B (150 mM NaCl, 100 mM sodium bicarbonate, pH 8.5) for 30 min. The cells were washed in buffer B and a cover slip was added to each slide, over a few drops of Vectashield (Vector Laboratories, Burlingame, Calif.). The slides were stored in the dark until ready for microscopic examination.  
     [0111] Apo E Western Blotting  
     [0112] Cells were treated with Media was concentrated 20-fold by centrifugal ultrafiltration (VIVA SPIN 20, MCO 5,000, Viva Sciences, Hannover, Germany), dialyzed against 0.15M NaCl, 1 mM sodium EDTA, 0.025% sodium azide (SalEN). Total protein content was determined by a modified Lowry assay (BioRad DC kit, Richmond, Calif.). Concentrated media (50 μg protein) was made to Start Buffer (0.025 M NaCl, 0.010 M tris (pH 8.5), 5 mM MnCl 2 ) and adsorbed onto a 0.1 ml column containing Heparin-Sepharose CL-4B (Pharmacia, Uppsala, Sweden). Following a 2 ml wash in Start Buffer, the apo E containing bound fraction was eluted with 0.5M NaCl in Start buffer. The eluate was concentrated to 20 μg and buffer-exchanged to SalEN by centrifugal ultrafiltration (Biomax, 5k MCO, Millipore, Bedford, Mass.). Apo E was resolved by tris-tricine buffered SDS-PAGE (5-25% linear acrylamide gradient) and proteins electrophoetically transferred (55V, 18 h) to nitrocelluose membrane filters (Schleicher and Shuell, Keen, N.H.). Membranes were blocked with 10% bovine serum albumin at room temperature and probed with 1% goat anti-human apo E antiserum (18 h, 3° C.) prepared in 0.15% NaCl, 1 mM EDTA (pH 7.4), 0.1% Triton X-100 (SalET). The primary-bound anti-apo E antibodies were detected colorimetrically with horseradish peroxidase conjugated rabbit anti-goat Ig (H+L) and NiCl 2 -enhanced diaminobenzine staining. Stained bands were compared densitometrically from the digitized scanned image (NIH Image, v.1.62). Anti apo E antibodies were obtained by hyper-immunization of goats with purified apo E or obtained from Assay Designs (A299, Ann Arbor, Mich.)  
     [0113] Lipoprotein fractions were prepared from conditioned media that was adjusted with solid KBr to a density of 1.21 g/ml. Samples were ultracentrifuged in a Beckman 42.2 Ti rotor at 40,000 rpm for 18 h at 10° C. The lipoprotein and lipoprotein-free fractions, the top and bottom 50 μl, respectively, were dialysed against SalEN prior to analysis.  
     [0114] [ 14 C] Docosohexanoic Acid (DHA) Labeled POS Uptake and Transport  
     [0115] Bovine outer photoreceptor outer segments (POS) were labeled by incubating for with Coenzyme A, ATP, Mg  2+ , and [ 14 C]-DHA. Cells grown on laminin coated Transwell plates were incubated with 12 μg/ml labeled POS in the apical chamber for 36 hours in medium containing 10% lipoprotein deficient fetal bovine serum. The basal medium was subjected to scintillation counting to determine the amount of [ 14 C] labeled lipids transported through the RPE cells.  
     [0116] Identification of Acceptors for Exported  14 C Lipids  
     [0117] Bovine outer photoreceptor outer segments (POS) were labeled by incubating for with Coenzyme A, ATP, Mg  2+ , and [ 14 C]-DHA. Cells grown on laminin coated Transwell plates were incubated with 12 μg/ml labeled POS in the apical chamber for 36 hours in medium containing 10% lipoprotein deficient fetal bovine serum. The basal chambers contained either 1 mg/ml human HDL, 1 mg/ml human LDL or 100% human plasma. The basal medium was collected and lipoproteins were repurified from by potassium bromide density gradient centrifugation at d=1.21 g/ml (Beckman 42.2 Ti rotor, 40,000 rpm, 18 h, 10° C.), dialyzed, and resolved by size in nondenaturing 0-35% PAGE. Gels were stained with coomassie blue R-250. Gel lanes were sectioned into thirty 2 mm slices that were digested (TS-1, Research Products International) and radioactivity quanitfied by liquid scintillation spectrometry.  
     Example 2  
     EXPRESSION OF TRANSPORTERS IN RPE CELLS  
     [0118] One skilled in the art recognizes that certain RCT components in cultured human RPE cells have been demonstrated (Mullins et al,. 2000; Anderson et al., 2001). Nuclear hormone receptors known to regulate expression of reverse cholesterol transport proteins are also expressed in cultured human RPE.  
     [0119] A skilled artisan recognizes that there is expression of TRs and RXRs in RPE cells in culture (Duncan et al. 1999). RT-PCR of human RPE cell cDNA revealed that these cells also express mRNAs for apo E, ABCA1, SR-BI, SR-BII and lxr α. As shown in FIG. 1 lane  1 , FIG. 1 lane  2  and FIG. 1 lane  3 , RPE cells express mRNAs for apo E, ABCA1 and lxr α, respectively.  
     [0120] As shown in FIG. 2, lane  1 , and FIG. 2, lane  2 , RPE cells express mRNA for SR-BI and SR-BII respectively.  
     [0121] Furthermore, in immunofluoresence microscopy experiments, RPE cells stain strongly for SR-BI (FIG. 3A) and SR-BII (FIG. 3B). Control non-specific IgG or antibody vehicle did not stain RPE cells (FIGS. 3C and 3D, respectively). Expression of SR-BI and SR-BII in these cells was confirmed by PCR.  
     [0122] Expression of ABCA1 protein was demonstrated by immunofluorescent staining of RPE cells with an antibody to ABCA1 (FIG. 4). Cell nuclei were stained with DAPI.  
     Example 3  
     REGULATION OF APO E SECRETION IN RPE CELLS  
     [0123] In order to distinguish apical (A) from basally (B) secreted apo E, RPE cells were cultured on laminin-coated Transwell plates. Specifically, human cultured RPE (passage  2 - 10 , 35 y.o. donor) were placed on laminin-coated Transwell plates, wherein the upper and lower wells both had serum-free media. Total protein and apo E-specific protein concentrations were measured from media pooled and concentrated from 3-6 replicate wells. To assess apo E-specific secretion, apo E was purified from conditioned media by heparin-sepharose affinity chromatography and visualized by western blotting. Apo E concentrations were consistently greater in the basolateral media (FIG. 5, lane  1  vs. lane  2 ). These data demonstrate that RPE cells display polarized secretion of cellular proteins, including apo E. Thus, this indicated that Apo E is preferentially secreted basally, supporting its role in RCT.  
     [0124] Since RPE cells express lxr α as well as thyroid hormone receptors (TRs) and retinoid-X-receptors (RXRs), the effect of 10 −7  M T3, 2.5×10 −6  M 22 (R) hydroxycholesterol (HC) (an lxr α agonist), or 10 −7  M cis retinoic acid (cRA) (an RXR agonist) on apo E secretion from RPE cells was tested. FIG. 6 illustrates the same experimental procedure as described above, but with basal and apical media both containing the following compounds for a 36 hour incubation: T3 (10 −7 ) M (T); 9 cis-RA (10 −6 ) M (RA); and 22 (R) hydroxycholesterol 2.5 (10 −6 ) M (HC). The basal media was analyzed for Apo E expression with Western blot, and the results showed increased basal expression of Apo E with the compound treatments. Thus, as before, polarized apo E secretion was observed and, in this case, occurred in the presence of T3, HC or cRA, indicating that an increase in levels of basally secreted apo E is the result of administration of these compounds to RPE cells.  
     Example 4  
     ASSAY OF EFFLUX FROM RPE CELLS  
     [0125] This example characterizes efflux of POS residues from RPE cells, particularly regarding binding to HDL. Giusto et al. (1986) describes a method of  14 C decoshexanoic acid (DHA) labeling of bovine photoreceptor outer segment (POS) lipids. Generally, an approximately 36 hour incubation over human RPE cells wherein the basal medium contains plasma, HDL, or LDL is followed by centrifugation of the basal media to collect lipoprotein fraction, which is then analyzed to determine distribution of radioactivity.  
     [0126] Specifically, bovine photoreceptor outer segment (POS) are labeled with  14 C decoshexanoic acid (DHA) and placed in lipoprotein deficient media. Following this, they are placed over cultured human RPE on Transwell plates for 36 hours, and the basal medium contained either 100% plasma, HDL (1 mg/cc) or LDL (1 mg/cc). After 36 hours, basal media was centrifuged to collect lipoprotein fraction (density 1.2). This fraction was then run on a non-denaturing gel and stained with Coomassie blue. FIG. 7 shows LDL and HDL fractions, both separately and together in plasma (PL). The PL fraction contains the same amount of HDL and LDL as each of the separated fractions (HDL, LDL).  
     [0127] The PA gel was cut into about 1 mm pieces, and the radioactivity distribution was determined (FIG. 8). With either LDL or HDL alone, counts were observed over respective lipoprotein fractions. When both LDL and HDL in plasma are present, counts localize preferentially over HDL fraction. This indicates that following phagocytosis of POS by RPE, POS residues are effluxed and preferentially bound by HDL. This is a novel demonstration illustrating that RCT to an HDL acceptor occurs in RPE cells.  
     [0128] To characterize the lipids in the lipoprotein fraction, thin layer chromatography was performed. Acid charring was used to identify lipid containing spots. The spots were scraped off of the plate and  14 C was quantified by liquid scintillation counting. Six of 17  14 C-containing spots were identified with standards shown (FIG. 9). Eleven  14 C-containing spots bound to HDL remain unidentified and could be unique serum marker(s) for patients with early AMD.  
     [0129] Thus, in an embodiment of the present invention, a patient sample is obtained, such as by drawing blood, and the HDL is examined for bound POS residues. From this, a determination of their risk of visual loss from AMD is made. In a specific embodiment, the profile of bound POS residues is indicative of identifying an individual afflicted with ocular disease and/or of identifying an individual at risk for developing an ocular disease.  
     Example 5  
     MODULATION OF RCT BY COMPOUND ADMINISTRATION  
     [0130] This experiment determines whether compound administration can upregulate efflux of labeled POS residues to HDL, particularly by showing regulation of  14 C-DHA labeled POS efflux into basal media. An assay similar to that described in Example 4 is utilized; however, in this Example the cells were treated with T3, 9 cis-retinoic acid, and 22 (R) hydroxycholesterol in the concentrations described above for 36 hours. Total radioactivity (cpm) in the absence of HDL purification was determined by liquid scintillation counting of the basal media. FIG. 10 indicates that compound treatments increase RCT by cultured human RPE cells.  
     [0131] Specifically, cells were grown for 1 to 2 weeks at confluence on Transwell plates.  14 C-labeled POS (30 mg/ml) were added to the apical medium. The apical and basal medium comprised either 10 −7  M T3, 2.5×10 −5  M 22 (R) hydroxycholesterol, or 10 −7  M cis retinoic acid. The basal medium contained 1 mg/ml HDL. After 36 hours the basal medium was collected and  14 C counts were determined by scintillation counting. As stated, all of the compound treatments increased transport of  14 C-labeled POS to the basal medium.  
     [0132] The effect of T3 on Apo E mRNA levels was also assessed by RT-PCR. Treatment with 10 −7  M T3 resulted in a 1.5 to 2-fold increase in apo E mRNA levels, suggesting that T3 is acting, at least in part, to increase apo E levels at the mRNA level. In specific embodiments, administration of 9 cis-retinoic acid and 22 (R) hydroxycholesterol similarly upregulates expression of apo E.  
     [0133] Thus, in a specific embodiment, RCT is regulated via nuclear hormone receptor ligands. For example, ABCA1 expression is upregulated by binding of LXR and RXR agonists to their respective nuclear hormone receptors (FIG. 11). Since these receptors form heterodimers bound to the ABCA1 promoter, ligand binding increases expression of ABCA1 and, hence, RCT.  
     Example 6  
     IDENTIFICATION OF HDL AS LIPID ACCEPTOR FROM RPE CELLS  
     [0134] In the presence of added purified human LDL and HDL, radiolabeled lipid efflux is enhanced (FIG. 12). As shown graphically, efflux (bottoms in graph) was greatly enhanced by the presence of plasma (PL in graph), HDL or LDL, as compared to no addition to the bottom medium (left side of graph).  
     [0135] As shown in FIG. 8, when whole human EDTA-plasma is employed and lipoproteins are isolated, [ 14 C]-labeled lipids are incorporated into LDL and HDL. However, radiolabel preferentially associated with HDL. Furthermore, the radiolabel in HDL was localized to the larger HDL 2 subspecies, which include the HDL particles enriched in apo E. This result suggests that lipid efflux from RPE is enhanced by the apo E—containing HDL.  
     Example 7  
     REDUCTION OF BM LIPIDS VIA SCAVENGER RECEPTORS (SRS)  
     [0136] Scavenger receptors in macrophages function to phagocytose oxLDL molecules. There are types of SRs previously described in macrophages including SR-A1, SR-A2, SR-B1, SR-B2, CD36, and LOX. SRs are distinct from LDL receptors in that levels of expression for SRs are upregulated by oxLDL. This upregulation by intracellular oxLDL levels is modulated by nuclear hormone receptors, peroxisome proliferator activated receptor (PPAR) and retinoic acid X receptor (RXR), that exert transcriptional control of CD36 expression. Because the earliest lesion of AS, the fatty streak, consists of macrophages engulfed with excessive oxLDL, and because RPE cells similarly become filled with lipid inclusions in AMD, SR expression was studied in RPE cells. Expression of the following SRs in RPE cells was identified: CD36 (confirmation of previous investigators), SR-A1, SR-A2 (both first time demonstrated in RPE), SR-B1, SR-B2 (both first time demonstrated in RPE).  
     [0137] The inventors have also shown that, like macrophages, oxLDL upregulates expression of CD36 in RPE cells (FIG. 13). Additionally, RPE cells express the nuclear hormone receptors, PPAR and RXR, indicating control mechanisms for SR expression are analogous between the cell types. Thus, in specific embodiments the expression of RPE SRs in patients is controlled with PPAR and RXR ligands (e.g. PG-J2, thiazolidinediones, cis-retinoic acid). This controls the rate at which RPE cells phagocytose oxidized photoreceptor outer segments, and hence slows the rate at which abnormal lipid inclusions accumulate in RPE and BM. In other specific embodiments, expression of CD36 is downregulated with a composition such as tamoxifen, TGF-beta or INF-gamma. Similarly, regulating expression of other RPE SRs would control levels of lipids in both RPE and BM. For example, for SR-A regulation IGF-1, TGF-beta, EGF, and/or PDGF is used, and for SR-B regulation cAMP and/or estradiol (for upregulation) or TNF-alpha, LPS, and/or INF-gamma (for downregulation) is used.  
     Example 8  
     HDL INCREASES  14 C LIPID EFFLUX FROM RPE CELLS PREFERENTIALLY TO OTHER LIPOPROTEINS  
     [0138] Transcription of the apo E gene is regulated by liver-X-receptor alpha (LXR α) that acts as heterodimers with retinoid-X-receptor alpha (RXR α) (Mak et al., 2002). The inventors have previously shown that RPE cells express T 3  receptors (TRs) that also act as heterodimers with RXR α (Duncan et al., 1999). The inventors and others, have demonstrated that primary cultures of RPE cells express mRNA for lxr α, RXR α, apo E, and other proteins involved in regulation of lipid and cholesterol uptake, metabolism and efflux (summarized herein). In this Example, the inventors show that apo E secreted by primary cultures of RPE cells can be up-regulated by thyroid hormone (T 3 ), 22(R) hydroxycholesterol (HC), and cis retinoic acid (RA). The inventors also demonstrated that a high density lipoprotein (HDL) fraction rich in apo E is a preferential acceptor for labeled POS lipids.  
     [0139] As shown in the Table, the present inventors and other investigators have identified mRNAs for the proteins involved in regulating lipid and cholesterol uptake, metabolism and efflux. The cells used in the experiments described below express only the apo E3 allele (E3/E3).  
                                          TRANSCRIPTION               FACTORS       LIGANDS               TR α1   Thyroid hormone Receptor alpha 1   T 3         TR α2   Thyroid hormone Receptor alpha 2   T 3         TR β1   Thyroid hormone Receptor beta 1   T 3         RXR α   Retinoid-X Receptor alpha   Retinoic               Acid       RXR α   Retinoid-X Receptor beta   Retinoic               Acid       PPAR α   Peroxisome Proliferator   Oxidized           Activator Receptor gamma   lipids       Lxr α   Liver-X Receptor alpha   Oxysterols               CELL SURFACE               RECEPTORS       LIGANDS               SR-BI   Scavenger Receptor BI   Oxidized               Lipids       SR-BII   Scavenger Receptor BII   Oxidized               Lipids       SR-AI   Scavenger Receptor AI   Oxidized               Lipoproteins       SR-AII   Scavenger Receptor AII   Oxidized               Lipoproteins       Lox-1   Lectin-like Oxidized LDL   Oxidized           receptor 1   Lipoproteins               CHOLESTEROL/               LIPID               TRANSPORT AND               METABOLISM       FUNCTIONS               SR-BI   Scavenger Receptor BI   Reverse               Cholesterol               Transport       SR-BII   Scavenger Receptor BII   Reverse               Cholesterol               Transport       ABCA1   ATP Binding Cassette   Reverse           Protein A1   Cholesterol               Transport       ACAT1   Acyl-CoA Cholesterol   Cholesterol           Acyltransferase 1   Acylation       Apo E   Apolipoprotein E   Cholesterol/               Lipid               Trafficking                  
 
     [0140] As shown qualitatively in FIG. 6, T 3  (TR agonist), RA (RXR agonist), HC (LXR agonist) stimulate basal apo E secretion. As previously indicated, RPE cells were treated for 36 hours on Transwell® plates with serum free DMEM in upper and lower chambers +/− the drugs indicated. Control (C ) refers to no drug addition; T refers to 10 −7  M T 3 ; RA refers to 10 −7  M cis retinoic acid; and HC refers to 2.5×10 −6  M 22(R) hydroxycholesterol. Basal media from 3 wells were combined, concentrated, and apo E was detected by western blotting.  
     [0141] As shown quantitatively in FIG. 14, TR, LXR and RXR agonists upregulate apo E secretion alone and in combination, as assessed by ELISA assays. RPE cells were treated for 36 hours on Transwell® plates with serum free DMEM +/− the drugs indicated. Control refers to no drug addition, T refers to 10 −7  M T 3 ; HC refers to 2.5×10 −6  M 22(R) hydroxycholesterol; RA refers to 10 −7  M cis retinoic acid. N=6, * indicates p≦0.05 (two-tailed t-test) compared to Control.  
     [0142] As shown in FIG. 15, apo E secreted from RPE cells binds to HDL. RPE cells on Transwell® plates were grown in DMEM with 5% FBS for 36 hours (apical chamber). Basal chambers had serum free DMEM with either 200, 50, or 0 μg/ml mouse HDL (lanes  2 ,  3 , and  4  respectively. Lane  1  illustrates molecular weight markers. HDL was purified by ultracentrifugation, resolved by polyacrylamide gel electrophoresis, and human apo E was identified by western blotting.  
     [0143] As shown in FIG. 16, HDL stimulates POS lipid efflux from RPE cells in culture. RPE cells on Transwell® plates were fed  14 C labeled POS in DMEM with 5% FBS for 36 hours (apical chamber). Basal chambers had serum free DMEM. Both upper and lower media contained either no addition (Control), 10% human plasma, 100 μg/ml HDL, 1000 μg/ml LDL or 50 μg/ml HDL+500 μg/ml LDL as indicated. FIG. 16 left: Basal  14 C cpm/130 μl. N=3, * indicates p≦0.05 (two-tailed t-test) compared to Control. FIG. 16 right: Lipoproteins were purified by ultracentrifugation, dialyzed to remove soluble  14 C, and counted.  
     [0144] As shown in FIG. 8,  14 C labeled POS lipids preferentially bind to apo E containing high molecular weight HDL (HDL3).  14 C labeled lipoproteins from the lower chamber were purified by ultracentrifugation and resolved on native polyacrylamide gels.  
     [0145] Characterization of HDL and plasma bound POS lipids was made by thin layer chromatography, as shown in FIG. 17.  14 C labeled lipoproteins from the lower chamber were purified by ultracentrifugation, and lipids were resolved by thin layer chromatography followed by acid charring.  
     [0146] As shown in FIG. 18, six spots in HDL and plasma were tentatively identified; at least 11 other spots are not yet identified. Spots identified by charring were cut out and  14 C cpm determined by liquid scintillation counting.  
     Example 9  
     EXEMPLARY METHODS AND MATERIALS FOR EXAMPLE 8  
     [0147] Cell Culture  
     [0148] Primary cultures of normal human RPE cells were prepared from a 35 year old donor eye as described (Song and Lui, 1990). Cells from passages 4 to 10 were used. RPE cells were grown on laminin-coated tissue culture plates, or on laminin coated 0.4 μM cellulose acetate Transwell® dishes (Costar) in DMEM H21 containing 5-10% fetal bovine serum (FBS), 2 mM glutamine, 5 μg/ml gentamycin, 100 IU/ml penicillin, 100 mg/ml streptomycin, 2.5 mg/ml fungizone, 1 ng/ml bFGF, and 1 ng/ml EGF. Where indicated, FBS was substituted with: 6 g/L NEAA, 0.39 g/L methylcellulose (serum free medium). No differences in cell morphology or protein expression were observed in cultures from different passages. RPE cells were grown at confluence for at least 14 days prior to undergoing the experimental treatments described below.  
     [0149] RT-PCR  
     [0150] RT-PCR was carried out on 1 μg of cDNA. The RT-PCR products are resolved by electrophoresis on 1.4% agarose gels. The RT-PCR primer sequences used are given followed by the predicted apo E RT-PCR product size. apo E forward: 5′- TAA GCT TGG CAC GGC TGT CCA AGG A (SEQ ID NO:13); apo E reverse: 5′-ACA GAA TTC GCC CCG GCC TGG TAC AC (SEQ ID NO:14); 241 bp product (detects both apo E3 and apo E4). PCR was conducted for 20-30 cycles at 55° C. in buffer containing 2.0-5.0 mM MgCl 2 . The RT-PCR product of the predicted size for apo E had its identity confirmed by DNA sequencing. Only the apo E3 mRNA sequence was detected. Messenger RNAs for the other proteins listed in the table were identified using similar strategies with primers specific to each cDNA.  
     [0151] Western Blotting  
     [0152] Briefly, media was concentrated 20-fold by centrifugal ultrafiltration (VIVA SPIN 20, MCO 5,000; Viva Sciences). Concentrated media (20 μg protein) was purified over Heparin-Sepharose CL-4B. The apo E containing (bound) fraction was eluted and re-concentrated to 20 μl. Apo E was resolved by 5-25% linear gradient SDS polyacrylamide gel electrophoresis, and proteins were electrophoretically transferred to nitrocelluose. Membranes were blocked with 10% BSA and probed with 1% goat anti-human apo E antiserum. Apo E antibodies were detected colorimetrically with horseradish peroxidase conjugated rabbit anti-goat IgG and NiCl 2 -enhanced diaminobenzine staining.  
     [0153] ELISA Assay  
     [0154] Media samples treated with 0.1% Tween-20 containing 1% bovine serum albumin were incubated (37° C., 4 h) in 96-well plates previously coated with apo E-affinity purified goat anti-apo E antibody. Apo E was detected using a secondary antibody-peroxidase conjugate and 3.3.5.5′-tetramethylethylenediamine (TMB) substrate. Optical density was measured at 450 nm. The assay was calibrated with purified plasma apo E. The dynamic range of the assay was 1-40 ng/ml apo E with a CV&lt;5%.  
     [0155] POS Lipid Transport and Lipoprotein Gel Analysis  
     [0156] Briefly, purified POS lipids were labeled with  14 C docosohexanoic acid as described (Guisto et al., 1986). Twenty μg/ml (protein) of POS were added to the top chambers of 6 well Transwell® plates. The bottom chambers contained serum free medium with or without human high density lipoprotein (HDL), human low density lipoprotein (LDL), or human plasma in the amounts indicated. After 36 hours, cell culture medium was harvested from the bottom chambers, adjusted to a density of 1.25 g/ml with solid potassium bromide and underlayed over a KBr solution of d=1.21. Samples were ultracentrifuged (Beckman 50.2Ti, 45,000 rpm, 10° C.) for 20 hours. The top (lipoprotein) layer was removed, dialyzed, and subjected to non-denaturing gel electrophoresis. The gels were stained with Coomassie Blue and photographed, after which 2 mM slices were subjected to scintillation counting.  
     [0157] Thin Layer Chromatography  
     [0158] Lipoprotein samples were extracted for lipid by the method of Bligh-Dyer, which is well known in the art. Lipids were resolved by silica gel K6 thin layer chromtography using sequential developments in Solvent 1: chloroform/methanol/acetic acid/water (25:15:4:2) and Solvent 2: n-hexane/diethylether/acetic acid (65:35:2). Lipid species were detected by acid charring, plates were immersed in 7.5% copper acetate, 2.5% copper sulfate, 8% phosphoric acid and heated on a hot plate for 1 hour.  14 C radioactivity was measured by liquid scintillation counting in standard methods known in the art.  
     REFERENCES  
     [0159] All patents and publications mentioned in the specification are indicative of the level of those skilled in the art to which the invention pertains. All patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. The references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference.  
     Patents  
     [0160] U.S. Pat. No. 6,071,924  
     [0161] U.S. Pat. No. 5,846,711  
     [0162] WO 00/52479  
     [0163] WO 01/58494  
     [0164] WO 02/13812  
     Publications  
     [0165] Anderson DH, Ozaki S, Nealon M, Neitz J, Mullins RF, Hageman GS, Johnson LV: Local cellular sources of apolipoprotein E in the human retina and retinal pigmented epithelium: implications for the process of drusen formation.:Am J Ophthalmol. 2001;131(6):767-8  
     [0166] Anderson, D. H. et al. (2001) Local Cellular Sources of Apolipoprotein E in the Human Retina and Retinal Pigmented Epithelium: Implications for the Process of Drusen Formation, Amer. J. Ophthalm. 131(6):767-781.  
     [0167] Bellosta S, Mahley R, Sanan D, Murata J, Newland D, Taylor J, Pitas R. Macrophage-specific expression of human apolipoprotein E reduces atherosclerosis in hypercholesterolemic E-null mice. J Clin Invest. 1995;96:2170-217  
     [0168] Browning PJ, Roberts DD, Zabrenetzky V, Bryant J, Kaplan M, Washington RH, Panet A, Gallo RC, Vogel T: Apolipoprotein E (ApoE), a novel heparin-binding protein inhibits the development of Kaposi&#39;s sarcoma-like lesions in BALB/c nu/nu mice. J Exp Med. 1994; 180(5): 1949-54  
     [0169] Curcio, C. A., K.Bradley, C.Guidry, M.Kirk, L.Wilson, S.Bames, H. S. Kruth, C. C. Y. Chang, T. Y. Chang: A Local Source for Esterified Cholesterol (EC) in Human Bruch&#39;s Membrane (BrM), ARVO Abstracts 2002 (need ref)  
     [0170] Curcio, Christine A., Millican, C. Leigh, Bailey, Tammy, Kruth, Howard S. Accumulation of Choslesterol with Age in Human Bruch&#39;s Membrane. Invest. Ophthalmol. Vis. Sci. 2001 42: 265-274  
     [0171] Dithmar, Stefan, Curcio, Christine A., Le, Ngoc-Anh, Brown, Stephanie, Grossniklaus, Hans E.:Ultrastructural Changes in Bruch&#39;s Membrane of Apolipoprotein E-Deficient Mice Invest. Ophthalmol. Vis. Sci. 2000 41: 2035-2042  
     [0172] Duncan KG, Bailey KR, Baxter JD, Schwartz DM. The human fetal retinal pigment epithelium: A target tissue for thyroid hormones. Ophthalmic Res. 1999; 31(6):399-406.  
     [0173] Feeney-Burns L, Hilderbrand ES, Eldridge S: Aging human RPE: morphometric analysis of macular, equatorial, and peripheral cells. Invest Ophthalmol Vis Sci 1984; 25: 195-200.  
     [0174] Friedman, E. (2000) The Role of the Atherosclerotic Process in the Pathogenesis of Age-related Macular Degeneration, Amer. J. Ophthalm. 130(5):658-663  
     [0175] Guisto NM, de Boschero MI, Sprecher H, Aveldano MI: Active labeling of phosphatidylcholines by [1-14C]docosahexaenoate in isolated photoreceptor membranes. Biochim Biophys Acta 1986;860:137-48  
     [0176] Hasty, A. H., M. F. Linton, S. J. Brandt, V. R. Babaev, L. A. Gleaves, and S. Fazio. 1999. Retroviral gene therapy in ApoE-deficient mice: ApoE expression in the artery wall reduces early foam cell lesion formation. Circulation. 99: 2571-2576  
     [0177] Holz FG, Sheraidah G, Pauleikhoff D, Bird AC: Analysis of lipid deposits extracted from human macular and peripheral Bruch&#39;s membrane. Arch Ophthalmol 1994; 112: 402-406.  
     [0178] Janowski BA, Grogan MJ, Jones SA, Wisely GB, Kliewer SA, COrey EJ, Mangelsdorf DJ: Structural requirements of ligands for the oxyserol liver X receptors LXRa and LXRb. Proc Natl Acad Sci (USA) 1999 January 96:266-271.  
     [0179] Kennedy CJ, Rakoczy PE, Constable IJ: Lipofuscin of the retinal pigment epithelium: a review. Eye 1995; 9:262-274.  
     [0180] Klaver CC, Kliffen M, van Duijn CM, Hofman A, Cruts M, Grobbee DE, van Broeckhoven C, de Jong PT: Genetic association of apolipoprotein E with age-related macular degeneration. Am J Hum Genet. 1998 July;63(l):200-6  
     [0181] Kliffen, Mike, Lutgens, Esther, Daemen, Mat J A P, de Muinck, Ebo D, Mooy, Cornelia M, de Jong, Paulus T V M: The APO*E3-Leiden mouse as an animal model for basal laminar deposit Br J Ophthalmol 2000 84: 1415-1419  
     [0182] Laffitte BA, Repa JJ, Joseph SB, Wilpitz DC, Kast HR, Mangelsdorf DJ, Tontonoz P: LXRs control lipid-inducible expression of the apolipoprotein E gene in macrophages and adipocytes. Proc Natl Acad Sci USA 2001 January 16;98(2):507-12  
     [0183] Laffitte BA, Repa JJ, Joseph SB, Wilpitz DC, Kast HR, Mangelsdorf DJ, Tontonoz P:The modulation of apolipoprotein E gene expression by 3,3′-5-triiodothyronine in HepG2 cells occurs at transcriptional and post-transcriptional levels. Eur J Biochem. 1994 September 1;224(2):463-71.  
     [0184] Langer C, Huang Y, Cullen P, Wiesenhütter B, Mahley RW, Assmann G, von Eckardstein A. Endogenous apolipoprotein E modulates cholesterol efflux and cholesterol ester hydrolysis mediated by high-density lipoprotein-3 and lipid-free apolipoproteins in mouse peritoneal macrophages. J Mol Med. 2000;78:217-227  
     [0185] Lin, C. Y., H. W. Duan, and T. Mazzone. 1999. Apolipoprotein E-dependent cholesterol efflux from macrophages: kinetic study and divergent mechanisms for endogenous versus exogenous apolipoprotein E. J. Lipid Res. 40: 1618-1626  
     [0186] Mak PA, Laffitte BA, Desrumaux C, Joseph SB, Curtiss LK, Mangelsdorf DJ, Tontonoz P, Edwards PA: Regulated expression of the apolipoprotein E/C-I/C-IV/C-II gene cluster in murine and human macrophages. A critical role for nuclear liver X receptors alpha and beta. J Biol Chem. 2002 August 30;277(35):31900-8  
     [0187] Malek G, Li CM, Guidry C, Medeiros NE, Curcio CA: Apolipoprotein B in cholesterol-containing drusen and Basal deposits of human eyes with age-related maculopathy. Am J Pathol 2003;162(2):413-25  
     [0188] Mazzone T, Reardon C. Expression of heterologous human apolipoprotein E by J774 macrophages enhances cholesterol efflux to HDL3. J Lipid Res. 1994;35:1345-1353  
     [0189] Mazzone, T., L. Pusteinikas, and C. Reardon. 1992. Secretion of apoE by macrophages is accompanied by enhanced cholesterol efflux. Circulation. 86 (Suppl. I): 1-2  
     [0190] Michael E. Kelly, Moira A. Clay, Meenakshi J. Mistry, Hsiu-Mei Hsieh-Li and Judith A. K. Harmony: Apolipoprotein E Inhibition of Proliferation of Mitogen-Activated T Lymphocytes: Production of Interleukin 2 with Reduced Biological Activity, Cellular Immunology, Volume 159, Issue 2, December 1994, Pages 124-139.  
     [0191] Moore DJ, Clover GM: The effect of age on the macromolecular permeability of human Bruch&#39;s membrane. Invest Ophthalmol Vis Sci 2001; 42: 2970-2975  
     [0192] Moore DJ, Hussain AA, Marshall J: Age-related variation in the hydraulic conductivity of Bruch&#39;s membrane. Invest Ophthalmol Vis Sci 1995; 36: 1290-1297.  
     [0193] Mullins RF, Russell SR, Anderson DH, Hageman GS. Drusen associated with aging and age-related macular degeneration contain proteins common to extracellular deposits associated with atherosclerosis, elastosis, amyloidosis, and dense deposit disease. FASEB J. 2000 May;14(7):835-46.  
     [0194] Pauleikhoff D, Harper CA, Marshall J, Bird AC: Aging changes in Bruch&#39;s membrane. A histochemical and morphologic study. Ophthalmology 1990; 97: 171-178.  
     [0195] Sergio Fazio, Vladimir R. Babaev, Alisa B. Murray, Alyssa H. Hasty, Kathy J. Carter, Linda A. Gleaves, James B. Atkinson, and MacRae F. Linton: Increased atherosclerosis in mice reconstituted with apolipoprotein E null macrophages PNAS 94: 4647-4652  
     [0196] Sheraidah G, Steinmetz R, Maguire J, Pauleikhoff D, Marshall J, Bird AC: Correlation between lipids extracted from Bruch&#39;s membrane and age. Ophthalmology 1993; 100:47-51.  
     [0197] Shimano, H., J. Ohsuga, M. Shimada, Y. Namba, T. Gotoda, K. Harada, M. Katsuki, Y. Yazaki, and N. Yamada. 1995. Inhibition of diet-induced atheroma formation in transgenic mice expressing apolipoprotein E in the arterial wall. J. Clin. Invest. 95: 469-476  
     [0198] Simonelli F, Margaglione M, Testa F, Cappucci G, Manitto MP, Brancato R, Rinaldi E: Apolipoprotein E Polymorphisms in Age-Related Macular Degeneration in an Italian Population. Ophthalmic Res 2001;33:325-328  
     [0199] Song MK, Lui GM: Propagation of fetal human RPE cells: preservation of original culture morphology after serial passage. J Cell Physiol 1990;143:196-203  
     [0200] Souied EH, Benlian P, Amouyel P, Feingold J, Lagarde JP, Munnich A, Kaplan J, Coscas G, Soubrane G.: The epsilon4 allele of the apolipoprotein E gene as a potential protective factor for exudative age-related macular degeneration. Am J Ophthalmol. 1998;125(3):353-9  
     [0201] Spaide RF, Ho-Spaide WC, Browne RW, Armstrong D: Characterization of peroxidized lipids in Bruch&#39;s membrane. Retina 1999; 19: 141-147.  
     [0202] Starita C, Hussain AA, Pagliarini S, Marshall J: Hydrodynamics of ageing Bruch&#39;s membrane: implications for macular disease. Exp Eye Res 1996; 62: 565-572.  
     [0203] Tangirala RK, Pratico D, FitzGerald GA, Chun S, Tsukamoto K, Maugeais C, Usher DC, Pure E, Rader DJ: Reduction of isoprostanes and regression of advanced atherosclerosis by apolipoprotein E. J Biol Chem. 2001 January 5;276(1):261-6  
     [0204] Taylor, Hugh R, Keeffe, Jill E: World blindness: a 21st century perspective. Br J Ophthalmol 2001 85: 261-266  
     [0205] VanNewkirk, Mylan R., Nanjan, Mukesh B., Wang, Jie Jin, Mitchell, Paul, Taylor, Hugh R., McCarty, Cathy A.: The prevalence of age-related maculopathy: The visual impairment project Ophthalmology 2000 107: 1593-1600  
     [0206] Although the present invention and its advantages have been described in detail, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims. Moreover, the scope of the present application is not intended to be limited to the particular embodiments of the process, machine, manufacture, composition of matter, means, methods and steps described in the specification. As one of ordinary skill in the art will readily appreciate from the disclosure of the present invention, processes, machines, manufacture, compositions of matter, means, methods, or steps, presently existing or later to be developed that perform substantially the same function or achieve substantially the same result as the corresponding embodiments described herein may be utilized according to the present invention. Accordingly, the appended claims are intended to include within their scope such processes, machines, manufacture, compositions of matter, means, methods, or steps.  
    
     
       
         1 
         
           
             12  
           
           
             1  
             1156  
             DNA  
             Human  
           
            1 

cgcagcggag gtgaaggacg tccttcccca ggagccgact ggccaatcac aggcaggaag     60 

atgaaggttc tgtgggctgc gttgctggtc acattcctgg caggatgcca ggccaaggtg    120 

gagcaagcgg tggagacaga gccggagccc gagctgcgcc agcagaccga gtggcagagc    180 

ggccagcgct gggaactggc actgggtcgc ttttgggatt acctgcgctg ggtgcagaca    240 

ctgtctgagc aggtgcagga ggagctgctc agctcccagg tcacccagga actgagggcg    300 

ctgatggacg agaccatgaa ggagttgaag gcctacaaat cggaactgga ggaacaactg    360 

accccggtgg cggaggagac gcgggcacgg ctgtccaagg agctgcaggc ggcgcaggcc    420 

cggctgggcg cggacatgga ggacgtgtgc ggccgcctgg tgcagtaccg cggcgaggtg    480 

caggccatgc tcggccagag caccgaggag ctgcgggtgc gcctcgcctc ccacctgcgc    540 

aagctgcgta agcggctcct ccgcgatgcc gatgacctgc agaagcgcct ggcagtgtac    600 

caggccgggg cccgcgaggg cgccgagcgc ggcctcagcg ccatccgcga gcgcctgggg    660 

cccctggtgg aacagggccg cgtgcgggcc gccactgtgg gctccctggc cggccagccg    720 

ctacaggagc gggcccaggc ctggggcgag cggctgcgcg cgcggatgga ggagatgggc    780 

agccggaccc gcgaccgcct ggacgaggtg aaggagcagg tggcggaggt gcgcgccaag    840 

ctggaggagc aggcccagca gatacgcctg caggccgagg ccttccaggc ccgcctcaag    900 

agctggttcg agcccctggt ggaagacatg cagcgccagt gggccgggct ggtggagaag    960 

gtgcaggctg ccgtgggcac cagcgccgcc cctgtgccca gcgacaatca ctgaacgccg   1020 

aagcctgcag ccatgcgacc ccacgccacc ccgtgcctcc tgcctccgcg cagcctgcag   1080 

cgggagaccc tgtccccgcc ccagccgtcc tcctggggtg gaccctagtt taataaagat   1140 

tcaccaagtt tcacgc                                                   1156 

 
           
             2  
             5515  
             DNA  
             Human  
           
            2 

ggaacttgat gctcagagag gacaagtcat ttgcccaagg tcacacagct ggcaactggc     60 

agacgagatt cacgccctgg caatttgact ccagaatcct aaccttaacc cagaagcacg    120 

gcttcaagcc ctggaaacca caatacctgt ggcagccagg gggaggtgct ggaatctcat    180 

ttcacatgtg gggagggggc tcctgtgctc aaggtcacaa ccaaagagga agctgtgatt    240 

aaaacccagg tcccatttgc aaagcctcga cttttagcag gtgcatcata ctgttcccac    300 

ccctcccatc ccacttctgt ccagccgcct agccccactt tctttttttt ctttttttga    360 

gacagtctcc ctcttgctga ggctggagtg cagtggcgag atctcggctc actgtaacct    420 

ccgcctcccg ggttcaagcg attctcctgc ctcagcctcc caagtagcta ggattacagg    480 

cgcccgccac cacgcctggc taacttttgt atttttagta gagatggggt ttcaccatgt    540 

tggccaggct ggtctcaaac tcctgacctt aagtgattcg cccactgtgg cctcccaaag    600 

tgctgggatt acaggcgtga gctaccgccc ccagcccctc ccatcccact tctgtccagc    660 

cccctagccc tactttcttt ctgggatcca ggagtccaga tccccagccc cctctccaga    720 

ttacattcat ccaggcacag gaaaggacag ggtcaggaaa ggaggactct gggcggcagc    780 

ctccacattc cccttccacg cttggccccc agaatggagg agggtgtctg tattactggg    840 

cgaggtgtcc tcccttcctg gggactgtgg ggggtggtca aaagacctct atgccccacc    900 

tccttcctcc ctctgccctg ctgtgcctgg ggcaggggga gaacagccca cctcgtgact    960 

gggctgccca gcccgcccta tccctggggg agggggcggg acagggggag ccctataatt   1020 

ggacaagtct gggatccttg agtcctactc agccccagcg gaggtgaagg acgtccttcc   1080 

ccaggagccg gtgagaagcg cagtcggggg cacggggatg agctcagggg cctctagaaa   1140 

gagctgggac cctgggaagc cctggcctcc aggtagtctc aggagagcta ctcggggtcg   1200 

ggcttgggga gaggaggagc gggggtgagg caagcagcag gggactggac ctgggaaggg   1260 

ctgggcagca gagacgaccc gacccgctag aaggtggggt ggggagagca gctggactgg   1320 

gatgtaagcc atagcaggac tccacgagtt gtcactatca ttatcgagca cctactgggt   1380 

gtccccagtg tcctcagatc tccataactg gggagccagg ggcagcgaca cggtagctag   1440 

ccgtcgattg gagaacttta aaatgaggac tgaattagct cataaatgga acacggcgct   1500 

taactgtgag gttggagctt agaatgtgaa gggagaatga ggaatgcgag actgggactg   1560 

agatggaacc ggcggtgggg agggggtggg gggatggaat ttgaaccccg ggagaggaag   1620 

atggaatttt ctatggaggc cgacctgggg atggggagat aagagaagac caggagggag   1680 

ttaaataggg aatgggttgg gggcggcttg gtaaatgtgc tgggattagg ctgttgcaga   1740 

taatgcaaca aggcttggaa ggctaacctg gggtgaggcc gggttggggg cgctgggggt   1800 

gggaggagtc ctcactggcg gttgattgac agtttctcct tccccagact ggccaatcac   1860 

aggcaggaag atgaaggttc tgtgggctgc gttgctggtc acattcctgg caggtatggg   1920 

ggcggggctt gctcggttcc ccccgctcct ccccctctca tcctcacctc aacctcctgg   1980 

ccccattcag acagaccctg ggccccctct tctgaggctt ctgtgctgct tcctggctct   2040 

gaacagcgat ttgacgctct ctgggcctcg gtttccccca tccttgagat aggagttaga   2100 

agttgttttg ttgttgttgt ttgttgttgt tgttttgttt ttttgagatg aagtctcgct   2160 

ctgtcgccca ggctggagtg cagtggcggg atctcggctc actgcaagct ccgcctccca   2220 

ggtccacgcc attctcctgc ctcagcctcc caagtagctg ggactacagg cacatgccac   2280 

cacacccgac taactttttt gtattttcag tagagacggg gtttcaccat gttggccagg   2340 

ctggtctgga actcctgacc tcaggtgatc tgcccgtttc gatctcccaa agtgctggga   2400 

ttacaggcgt gagccaccgc acctggctgg gagttagagg tttctaatgc attgcaggca   2460 

gatagtgaat accagacacg gggcagctgt gatctttatt ctccatcacc cccacacagc   2520 

cctgcctggg gcacacaagg acactcaata catgcttttc cgctgggccg gtggctcacc   2580 

cctgtaatcc cagcactttg ggaggccaag gtgggaggat cacttgagcc caggagttca   2640 

acaccagcct gggcaacata gtgagaccct gtctctacta aaaatacaaa aattagccag   2700 

gcatggtgcc acacacctgt gctctcagct actcaggagg ctgaggcagg aggatcgctt   2760 

gagcccagaa ggtcaaggtt gcagtgaacc atgttcaggc cgctgcactc cagcctgggt   2820 

gacagagcaa gaccctgttt ataaatacat aatgctttcc aagtgattaa accgactccc   2880 

ccctcaccct gcccaccatg gctccaaaga agcatttgtg gagcaccttc tgtgtgcccc   2940 

taggtagcta gatgcctgga cggggtcaga aggaccctga cccgaccttg aacttgttcc   3000 

acacaggatg ccaggccaag gtggagcaag cggtggagac agagccggag cccgagctgc   3060 

gccagcagac cgagtggcag agcggccagc gctgggaact ggcactgggt cgcttttggg   3120 

attacctgcg ctgggtgcag acactgtctg agcaggtgca ggaggagctg ctcagctccc   3180 

aggtcaccca ggaactgagg tgagtgtccc catcctggcc cttgaccctc ctggtgggcg   3240 

gctatacctc cccaggtcca ggtttcattc tgcccctgtc gctaagtctt ggggggcctg   3300 

ggtctctgct ggttctagct tcctcttccc atttctgact cctggcttta gctctctgga   3360 

attctctctc tcagctttgt ctctctctct tcccttctga ctcagtctct cacactcgtc   3420 

ctggctctgt ctctgtcctt ccctagctct tttatataga gacagagaga tggggtctca   3480 

ctgtgttgcc caggctggtc ttgaacttct gggctcaagc gatcctcccg cctcggcctc   3540 

ccaaagtgct gggattagag gcatgagcac cttgcccggc ctcctagctc cttcttcgtc   3600 

tctgcctctg ccctctgcat ctgctctctg catctgtctc tgtctccttc tctcggcctc   3660 

tgccccgttc cttctctccc tcttgggtct ctctggctca tccccatctc gcccgcccca   3720 

tcccagccct tctcccccgc ctccccactg tgcgacaccc tcccgccctc tcggccgcag   3780 

ggcgctgatg gacgagacca tgaaggagtt gaaggcctac aaatcggaac tggaggaaca   3840 

actgaccccg gtggcggagg agacgcgggc acggctgtcc aaggagctgc aggcggcgca   3900 

ggcccggctg ggcgcggaca tggaggacgt gcgcggccgc ctggtgcagt accgcggcga   3960 

ggtgcaggcc atgctcggcc agagcaccga ggagctgcgg gtgcgcctcg cctcccacct   4020 

gcgcaagctg cgtaagcggc tcctccgcga tgccgatgac ctgcagaagc gcctggcagt   4080 

gtaccaggcc ggggcccgcg agggcgccga gcgcggcctc agcgccatcc gcgagcgcct   4140 

ggggcccctg gtggaacagg gccgcgtgcg ggccgccact gtgggctccc tggccggcca   4200 

gccgctacag gagcgggccc aggcctgggg cgagcggctg cgcgcgcgga tggaggagat   4260 

gggcagccgg acccgcgacc gcctggacga ggtgaaggag caggtggcgg aggtgcgcgc   4320 

caagctggag gagcaggccc agcagatacg cctgcaggcc gaggccttcc aggcccgcct   4380 

caagagctgg ttcgagcccc tggtggaaga catgcagcgc cagtgggccg ggctggtgga   4440 

gaaggtgcag gctgccgtgg gcaccagcgc cgcccctgtg cccagcgaca atcactgaac   4500 

gccgaagcct gcagccatgc gaccccacgc caccccgtgc ctcctgcctc cgcgcagcct   4560 

gcagcgggag accctgtccc cgccccagcc gtcctcctgg ggtggaccct agtttaataa   4620 

agattcacca agtttcacgc atctgctggc ctccccctgt gatttcctct aagccccagc   4680 

ctcagtttct ctttctgccc acatactgcc acacaattct cagccccctc ctctccatct   4740 

gtgtctgtgt gtatctttct ctctgccctt tttttttttt tagacggagt ctggctctgt   4800 

cacccaggct agagtgcagt ggcacgatct tggctcactg caacctctgc ctcttgggtt   4860 

caagcgattc tgctgcctca gtagctggga ttacaggctc acaccaccac acccggctaa   4920 

tttttgtatt tttagtagag acgagctttc accatgttgg ccaggcaggt ctcaaactcc   4980 

tgaccaagtg atccacccgc cggcctccca aagtgctgag attacaggcc tgagccacca   5040 

tgcccggcct ctgcccctct ttctttttta gggggcaggg aaaggtctca ccctgtcacc   5100 

cgccatcaca gctcactgca gcctccacct cctggactca agtgataagt gatcctcccg   5160 

cctcagcctt tccagtagct gagactacag gcgcatacca ctaggattaa tttggggggg   5220 

ggtggtgtgt gtggagatgg ggtctggctt tgttggccag gctgatgtgg aattcctggg   5280 

ctcaagcgat actcccacct tggcctcctg agtagctgag actactggct agcaccacca   5340 

cacccagctt tttattatta tttgtagaga caaggtctca atatgttgcc caggctagtc   5400 

tcaaacccct ggctcaagag atcctccgcc atcggcctcc caaagtgctg ggattccagg   5460 

catgggctcc gagcggcctg cccaacttaa taatattgtt cctagagttg cactc        5515 

 
           
             3  
             1157  
             DNA  
             Human  
           
            3 

ccccagcgga ggtgaaggac gtccttcccc aggagccgac tggccaatca caggcaggaa     60 

gatgaaggtt ctgtgggctg cgttgctggt cacattcctg gcaggatgcc aggccaaggt    120 

ggagcaagcg gtggagacag agccggagcc cgagctgcgc cagcagaccg agtggcagag    180 

cggccagcgc tgggaactgg cactgggtcg cttttgggat tacctgcgct gggtgcagac    240 

actgtctgag caggtgcagg aggagctgct cagctcccaa gtcacccaag aactgagggc    300 

gctgatggac gagaccatga aggagttgaa ggcctacaaa tcggaactgg aggaacaact    360 

gaccccggta gcggaggaga cgcgggcacg gctgtccaag gagctgcaga cggcgcaggc    420 

ccggctgggc gcggacatgg aggacgtgtg cggccgcctg gtgcagtacc gcggcgaggt    480 

gcaggccatg ctcggccaga gcaccgagga gctgcgggtg cgcctcgcct cccacctgcg    540 

caagctgcgt aagcggctcc tccgcgatcc cgatgacctg cagaagcgcc tggcagtgta    600 

ccaggccggg gcccgcgagg gcgccgagcg cggcctcagc gccatccgcg agcgcctggg    660 

gcccctggtg gaacagggcc gcgtgcgggc cgccactgtg ggctccctgg ccggccagcc    720 

gctacaggag cgggcccagg cctggggcga gcggctgcgc gcgcggatgg aggagatggg    780 

cagtcggacc cgcgaccgcc tggacgaggt gaaggagcag gtggcggagg tgcgcgccaa    840 

gctggaggag caggcccagc agatacgcct gcaggccgag gccttccagg cccgcctcaa    900 

gagctggttc gagcccctgg tggaagacat gcagcgccag tgggccgggc tggtggagaa    960 

ggtgcaggct gccgtgggca ccagcgccgc ccctgtgccc agcgacaatc actgaacgcc   1020 

gaagcctgca gccatgcgac cccacgccac cccgtgcctc ctgcctccgc gcagcctgca   1080 

gcgggagacc ctgtccccgc cccagccgtc ctcctggggt ggaccctagt ttaataaaga   1140 

ttcaccaagt ttcacgc                                                  1157 

 
           
             4  
             317  
             PRT  
             Human  
           
            4 

Met Lys Val Leu Trp Ala Ala Leu Leu Val Thr Phe Leu Ala Gly Cys 
1               5                   10                  15 

Gln Ala Lys Val Glu Gln Ala Val Glu Thr Glu Pro Glu Pro Glu Leu 
            20                  25                  30 

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

Gly Arg Phe Trp Asp Tyr Leu Arg Trp Val Gln Thr Leu Ser Glu Gln 
    50                  55                  60 

Val Gln Glu Glu Leu Leu Ser Ser Gln Val Thr Gln Glu Leu Arg Ala 
65                  70                  75                  80 

Leu Met Asp Glu Thr Met Lys Glu Leu Lys Ala Tyr Lys Ser Glu Leu 
                85                  90                  95 

Glu Glu Gln Leu Thr Pro Val Ala Glu Glu Thr Arg Ala Arg Leu Ser 
            100                 105                 110 

Lys Glu Leu Gln Ala Ala Gln Ala Arg Leu Gly Ala Asp Met Glu Asp 
        115                 120                 125 

Val Cys Gly Arg Leu Val Gln Tyr Arg Gly Glu Val Gln Ala Met Leu 
    130                 135                 140 

Gly Gln Ser Thr Glu Glu Leu Arg Val Arg Leu Ala Ser His Leu Arg 
145                 150                 155                 160 

Lys Leu Arg Lys Arg Leu Leu Arg Asp Ala Asp Asp Leu Gln Lys Arg 
                165                 170                 175 

Leu Ala Val Tyr Gln Ala Gly Ala Arg Glu Gly Ala Glu Arg Gly Leu 
            180                 185                 190 

Ser Ala Ile Arg Glu Arg Leu Gly Pro Leu Val Glu Gln Gly Arg Val 
        195                 200                 205 

Arg Ala Ala Thr Val Gly Ser Leu Ala Gly Gln Pro Leu Gln Glu Arg 
    210                 215                 220 

Ala Gln Ala Trp Gly Glu Arg Leu Arg Ala Arg Met Glu Glu Met Gly 
225                 230                 235                 240 

Ser Arg Thr Arg Asp Arg Leu Asp Glu Val Lys Glu Gln Val Ala Glu 
                245                 250                 255 

Val Arg Ala Lys Leu Glu Glu Gln Ala Gln Gln Ile Arg Leu Gln Ala 
            260                 265                 270 

Glu Ala Phe Gln Ala Arg Leu Lys Ser Trp Phe Glu Pro Leu Val Glu 
        275                 280                 285 

Asp Met Gln Arg Gln Trp Ala Gly Leu Val Glu Lys Val Gln Ala Ala 
    290                 295                 300 

Val Gly Thr Ser Ala Ala Pro Val Pro Ser Asp Asn His 
305                 310                 315 

 
           
             5  
             317  
             PRT  
             Human  
           
            5 

Met Lys Val Leu Trp Ala Ala Leu Leu Val Thr Phe Leu Ala Gly Cys 
1               5                   10                  15 

Gln Ala Lys Val Glu Gln Ala Val Glu Thr Glu Pro Glu Pro Glu Leu 
            20                  25                  30 

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

Gly Arg Phe Trp Asp Tyr Leu Arg Trp Val Gln Thr Leu Ser Glu Gln 
    50                  55                  60 

Val Gln Glu Glu Leu Leu Ser Ser Gln Val Thr Gln Glu Leu Arg Ala 
65                  70                  75                  80 

Leu Met Asp Glu Thr Met Lys Glu Leu Lys Ala Tyr Lys Ser Glu Leu 
                85                  90                  95 

Glu Glu Gln Leu Thr Pro Val Ala Glu Glu Thr Arg Ala Arg Leu Ser 
            100                 105                 110 

Lys Glu Leu Gln Ala Ala Gln Ala Arg Leu Gly Ala Asp Met Glu Asp 
        115                 120                 125 

Val Arg Gly Arg Leu Val Gln Tyr Arg Gly Glu Val Gln Ala Met Leu 
    130                 135                 140 

Gly Gln Ser Thr Glu Glu Leu Arg Val Arg Leu Ala Ser His Leu Arg 
145                 150                 155                 160 

Lys Leu Arg Lys Arg Leu Leu Arg Asp Ala Asp Asp Leu Gln Lys Arg 
                165                 170                 175 

Leu Ala Val Tyr Gln Ala Gly Ala Arg Glu Gly Ala Glu Arg Gly Leu 
            180                 185                 190 

Ser Ala Ile Arg Glu Arg Leu Gly Pro Leu Val Glu Gln Gly Arg Val 
        195                 200                 205 

Arg Ala Ala Thr Val Gly Ser Leu Ala Gly Gln Pro Leu Gln Glu Arg 
    210                 215                 220 

Ala Gln Ala Trp Gly Glu Arg Leu Arg Ala Arg Met Glu Glu Met Gly 
225                 230                 235                 240 

Ser Arg Thr Arg Asp Arg Leu Asp Glu Val Lys Glu Gln Val Ala Glu 
                245                 250                 255 

Val Arg Ala Lys Leu Glu Glu Gln Ala Gln Gln Ile Arg Leu Gln Ala 
            260                 265                 270 

Glu Ala Phe Gln Ala Arg Leu Lys Ser Trp Phe Glu Pro Leu Val Glu 
        275                 280                 285 

Asp Met Gln Arg Gln Trp Ala Gly Leu Val Glu Lys Val Gln Ala Ala 
    290                 295                 300 

Val Gly Thr Ser Ala Ala Pro Val Pro Ser Asp Asn His 
305                 310                 315 

 
           
             6  
             317  
             PRT  
             Human  
           
            6 

Met Lys Val Leu Trp Ala Ala Leu Leu Val Thr Phe Leu Ala Gly Cys 
1               5                   10                  15 

Gln Ala Lys Val Glu Gln Ala Val Glu Thr Glu Pro Glu Pro Glu Leu 
            20                  25                  30 

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

Gly Arg Phe Trp Asp Tyr Leu Arg Trp Val Gln Thr Leu Ser Glu Gln 
    50                  55                  60 

Val Gln Glu Glu Leu Leu Ser Ser Gln Val Thr Gln Glu Leu Arg Ala 
65                  70                  75                  80 

Leu Met Asp Glu Thr Met Lys Glu Leu Lys Ala Tyr Lys Ser Glu Leu 
                85                  90                  95 

Glu Glu Gln Leu Thr Pro Val Ala Glu Glu Thr Arg Ala Arg Leu Ser 
            100                 105                 110 

Lys Glu Leu Gln Thr Ala Gln Ala Arg Leu Gly Ala Asp Met Glu Asp 
        115                 120                 125 

Val Cys Gly Arg Leu Val Gln Tyr Arg Gly Glu Val Gln Ala Met Leu 
    130                 135                 140 

Gly Gln Ser Thr Glu Glu Leu Arg Val Arg Leu Ala Ser His Leu Arg 
145                 150                 155                 160 

Lys Leu Arg Lys Arg Leu Leu Arg Asp Pro Asp Asp Leu Gln Lys Arg 
                165                 170                 175 

Leu Ala Val Tyr Gln Ala Gly Ala Arg Glu Gly Ala Glu Arg Gly Leu 
            180                 185                 190 

Ser Ala Ile Arg Glu Arg Leu Gly Pro Leu Val Glu Gln Gly Arg Val 
        195                 200                 205 

Arg Ala Ala Thr Val Gly Ser Leu Ala Gly Gln Pro Leu Gln Glu Arg 
    210                 215                 220 

Ala Gln Ala Trp Gly Glu Arg Leu Arg Ala Arg Met Glu Glu Met Gly 
225                 230                 235                 240 

Ser Arg Thr Arg Asp Arg Leu Asp Glu Val Lys Glu Gln Val Ala Glu 
                245                 250                 255 

Val Arg Ala Lys Leu Glu Glu Gln Ala Gln Gln Ile Arg Leu Gln Ala 
            260                 265                 270 

Glu Ala Phe Gln Ala Arg Leu Lys Ser Trp Phe Glu Pro Leu Val Glu 
        275                 280                 285 

Asp Met Gln Arg Gln Trp Ala Gly Leu Val Glu Lys Val Gln Ala Ala 
    290                 295                 300 

Val Gly Thr Ser Ala Ala Pro Val Pro Ser Asp Asn His 
305                 310                 315 

 
           
             7  
             10412  
             DNA  
             Human  
           
            7 

gtaattgcga gcgagagtga gtggggccgg gacccgcaga gccgagccga cccttctctc     60 

ccgggctgcg gcagggcagg gcggggagct ccgcgcacca acagagccgg ttctcagggc    120 

gctttgctcc ttgttttttc cccggttctg ttttctcccc ttctccggaa ggcttgtcaa    180 

ggggtaggag aaagagacgc aaacacaaaa gtggaaaaca gttaatgacc agccacggcg    240 

tccctgctgt gagctctggc cgctgccttc cagggctccc gagccacacg ctgggggtgc    300 

tggctgaggg aacatggctt gttggcctca gctgaggttg ctgctgtgga agaacctcac    360 

tttcagaaga agacaaacat gtcagctgct gctggaagtg gcctggcctc tatttatctt    420 

cctgatcctg atctctgttc ggctgagcta cccaccctat gaacaacatg aatgccattt    480 

tccaaataaa gccatgccct ctgcaggaac acttccttgg gttcagggga ttatctgtaa    540 

tgccaacaac ccctgtttcc gttacccgac tcctggggag gctcccggag ttgttggaaa    600 

ctttaacaaa tccattgtgg ctcgcctgtt ctcagatgct cggaggcttc ttttatacag    660 

ccagaaagac accagcatga aggacatgcg caaagttctg agaacattac agcagatcaa    720 

gaaatccagc tcaaacttga agcttcaaga tttcctggtg gacaatgaaa ccttctctgg    780 

gttcctgtat cacaacctct ctctcccaaa gtctactgtg gacaagatgc tgagggctga    840 

tgtcattctc cacaaggtat ttttgcaagg ctaccagtta catttgacaa gtctgtgcaa    900 

tggatcaaaa tcagaagaga tgattcaact tggtgaccaa gaagtttctg agctttgtgg    960 

cctaccaagg gagaaactgg ctgcagcaga gcgagtactt cgttccaaca tggacatcct   1020 

gaagccaatc ctgagaacac taaactctac atctcccttc ccgagcaagg agctggctga   1080 

agccacaaaa acattgctgc atagtcttgg gactctggcc caggagctgt tcagcatgag   1140 

aagctggagt gacatgcgac aggaggtgat gtttctgacc aatgtgaaca gctccagctc   1200 

ctccacccaa atctaccagg ctgtgtctcg tattgtctgc gggcatcccg agggaggggg   1260 

gctgaagatc aagtctctca actggtatga ggacaacaac tacaaagccc tctttggagg   1320 

caatggcact gaggaagatg ctgaaacctt ctatgacaac tctacaactc cttactgcaa   1380 

tgatttgatg aagaatttgg agtctagtcc tctttcccgc attatctgga aagctctgaa   1440 

gccgctgctc gttgggaaga tcctgtatac acctgacact ccagccacaa ggcaggtcat   1500 

ggctgaggtg aacaagacct tccaggaact ggctgtgttc catgatctgg aaggcatgtg   1560 

ggaggaactc agccccaaga tctggacctt catggagaac agccaagaaa tggaccttgt   1620 

ccggatgctg ttggacagca gggacaatga ccacttttgg gaacagcagt tggatggctt   1680 

agattggaca gcccaagaca tcgtggcgtt tttggccaag cacccagagg atgtccagtc   1740 

cagtaatggt tctgtgtaca cctggagaga agctttcaac gagactaacc aggcaatccg   1800 

gaccatatct cgcttcatgg agtgtgtcaa cctgaacaag ctagaaccca tagcaacaga   1860 

agtctggctc atcaacaagt ccatggagct gctggatgag aggaagttct gggctggtat   1920 

tgtgttcact ggaattactc caggcagcat tgagctgccc catcatgtca agtacaagat   1980 

ccgaatggac attgacaatg tggagaggac aaataaaatc aaggatgggt actgggaccc   2040 

tggtcctcga gctgacccct ttgaggacat gcggtacgtc tgggggggct tcgcctactt   2100 

gcaggatgtg gtggagcagg caatcatcag ggtgctgacg ggcaccgaga agaaaactgg   2160 

tgtctatatg caacagatgc cctatccctg ttacgttgat gacatctttc tgcgggtgat   2220 

gagccggtca atgcccctct tcatgacgct ggcctggatt tactcagtgg ctgtgatcat   2280 

caagggcatc gtgtatgaga aggaggcacg gctgaaagag accatgcgga tcatgggcct   2340 

ggacaacagc atcctctggt ttagctggtt cattagtagc ctcattcctc ttcttgtgag   2400 

cgctggcctg ctagtggtca tcctgaagtt aggaaacctg ctgccctaca gtgatcccag   2460 

cgtggtgttt gtcttcctgt ccgtgtttgc tgtggtgaca atcctgcagt gcttcctgat   2520 

tagcacactc ttctccagag ccaacctggc agcagcctgt gggggcatca tctacttcac   2580 

gctgtacctg ccctacgtcc tgtgtgtggc atggcaggac tacgtgggct tcacactcaa   2640 

gatcttcgct agcctgctgt ctcctgtggc ttttgggttt ggctgtgagt actttgccct   2700 

ttttgaggag cagggcattg gagtgcagtg ggacaacctg tttgagagtc ctgtggagga   2760 

agatggcttc aatctcacca cttcggtctc catgatgctg tttgacacct tcctctatgg   2820 

ggtgatgacc tggtacattg aggctgtctt tccaggccag tacggaattc ccaggccctg   2880 

gtattttcct tgcaccaagt cctactggtt tggcgaggaa agtgatgaga agagccaccc   2940 

tggttccaac cagaagagaa tatcagaaat ctgcatggag gaggaaccca cccacttgaa   3000 

gctgggcgtg tccattcaga acctggtaaa agtctaccga gatgggatga aggtggctgt   3060 

cgatggcctg gcactgaatt tttatgaggg ccagatcacc tccttcctgg gccacaatgg   3120 

agcggggaag acgaccacca tgtcaatcct gaccgggttg ttccccccga cctcgggcac   3180 

cgcctacatc ctgggaaaag acattcgctc tgagatgagc accatccggc agaacctggg   3240 

ggtctgtccc cagcataacg tgctgtttga catgctgact gtcgaagaac acatctggtt   3300 

ctatgcccgc ttgaaagggc tctctgagaa gcacgtgaag gcggagatgg agcagatggc   3360 

cctggatgtt ggtttgccat caagcaagct gaaaagcaaa acaagccagc tgtcaggtgg   3420 

aatgcagaga aagctatctg tggccttggc ctttgtcggg ggatctaagg ttgtcattct   3480 

ggatgaaccc acagctggtg tggaccctta ctcccgcagg ggaatatggg agctgctgct   3540 

gaaataccga caaggccgca ccattattct ctctacacac cacatggatg aagcggacgt   3600 

cctgggggac aggattgcca tcatctccca tgggaagctg tgctgtgtgg gctcctccct   3660 

gtttctgaag aaccagctgg gaacaggcta ctacctgacc ttggtcaaga aagatgtgga   3720 

atcctccctc agttcctgca gaaacagtag tagcactgtg tcatacctga aaaaggagga   3780 

cagtgtttct cagagcagtt ctgatgctgg cctgggcagc gaccatgaga gtgacacgct   3840 

gaccatcgat gtctctgcta tctccaacct catcaggaag catgtgtctg aagcccggct   3900 

ggtggaagac atagggcatg agctgaccta tgtgctgcca tatgaagctg ctaaggaggg   3960 

agcctttgtg gaactctttc atgagattga tgaccggctc tcagacctgg gcatttctag   4020 

ttatggcatc tcagagacga ccctggaaga aatattcctc aaggtggccg aagagagtgg   4080 

ggtggatgct gagacctcag atggtacctt gccagcaaga cgaaacaggc gggccttcgg   4140 

ggacaagcag agctgtcttc gcccgttcac tgaagatgat gctgctgatc caaatgattc   4200 

tgacatagac ccagaatcca gagagacaga cttgctcagt gggatggatg gcaaagggtc   4260 

ctaccaggtg aaaggctgga aacttacaca gcaacagttt gtggcccttt tgtggaagag   4320 

actgctaatt gccagacgga gtcggaaagg attttttgct cagattgtct tgccagctgt   4380 

gtttgtctgc attgcccttg tgttcagcct gatcgtgcca ccctttggca agtaccccag   4440 

cctggaactt cagccctgga tgtacaacga acagtacaca tttgtcagca atgatgctcc   4500 

tgaggacacg ggaaccctgg aactcttaaa cgccctcacc aaagaccctg gcttcgggac   4560 

ccgctgtatg gaaggaaacc caatcccaga cacgccctgc caggcagggg aggaagagtg   4620 

gaccactgcc ccagttcccc agaccatcat ggacctcttc cagaatggga actggacaat   4680 

gcagaaccct tcacctgcat gccagtgtag cagcgacaaa atcaagaaga tgctgcctgt   4740 

gtgtccccca ggggcagggg ggctgcctcc tccacaaaga aaacaaaaca ctgcagatat   4800 

ccttcaggac ctgacaggaa gaaacatttc ggattatctg gtgaagacgt atgtgcagat   4860 

catagccaaa agcttaaaga acaagatctg ggtgaatgag tttaggtatg gcggcttttc   4920 

cctgggtgtc agtaatactc aagcacttcc tccgagtcaa gaagttaatg atgccatcaa   4980 

acaaatgaag aaacacctaa agctggccaa ggacagttct gcagatcgat ttctcaacag   5040 

cttgggaaga tttatgacag gactggacac caaaaataat gtcaaggtgt ggttcaataa   5100 

caagggctgg catgcaatca gctctttcct gaatgtcatc aacaatgcca ttctccgggc   5160 

caacctgcaa aagggagaga accctagcca ttatggaatt actgctttca atcatcccct   5220 

gaatctcacc aagcagcagc tctcagaggt ggctctgatg accacatcag tggatgtcct   5280 

tgtgtccatc tgtgtcatct ttgcaatgtc cttcgtccca gccagctttg tcgtattcct   5340 

gatccaggag cgggtcagca aagcaaaaca cctgcagttc atcagtggag tgaagcctgt   5400 

catctactgg ctctctaatt ttgtctggga tatgtgcaat tacgttgtcc ctgccacact   5460 

ggtcattatc atcttcatct gcttccagca gaagtcctat gtgtcctcca ccaatctgcc   5520 

tgtgctagcc cttctacttt tgctgtatgg gtggtcaatc acacctctca tgtacccagc   5580 

ctcctttgtg ttcaagatcc ccagcacagc ctatgtggtg ctcaccagcg tgaacctctt   5640 

cattggcatt aatggcagcg tggccacctt tgtgctggag ctgttcaccg acaataagct   5700 

gaataatatc aatgatatcc tgaagtccgt gttcttgatc ttcccacatt tttgcctggg   5760 

acgagggctc atcgacatgg tgaaaaacca ggcaatggct gatgccctgg aaaggtttgg   5820 

ggagaatcgc tttgtgtcac cattatcttg ggacttggtg ggacgaaacc tcttcgccat   5880 

ggccgtggaa ggggtggtgt tcttcctcat tactgttctg atccagtaca gattcttcat   5940 

caggcccaga cctgtaaatg caaagctatc tcctctgaat gatgaagatg aagatgtgag   6000 

gcgggaaaga cagagaattc ttgatggtgg aggccagaat gacatcttag aaatcaagga   6060 

gttgacgaag atatatagaa ggaagcggaa gcctgctgtt gacaggattt gcgtgggcat   6120 

tcctcctggt gagtgctttg ggctcctggg agttaatggg gctggaaaat catcaacttt   6180 

caagatgtta acaggagata ccactgttac cagaggagat gctttcctta acaaaaatag   6240 

tatcttatca aacatccatg aagtacatca gaacatgggc tactgccctc agtttgatgc   6300 

catcacagag ctgttgactg ggagagaaca cgtggagttc tttgcccttt tgagaggagt   6360 

cccagagaaa gaagttggca aggttggtga gtgggcgatt cggaaactgg gcctcgtgaa   6420 

gtatggagaa aaatatgctg gtaactatag tggaggcaac aaacgcaagc tctctacagc   6480 

catggctttg atcggcgggc ctcctgtggt gtttctggat gaacccacca caggcatgga   6540 

tcccaaagcc cggcggttct tgtggaattg tgccctaagt gttgtcaagg aggggagatc   6600 

agtagtgctt acatctcata gtatggaaga atgtgaagct ctttgcacta ggatggcaat   6660 

catggtcaat ggaaggttca ggtgccttgg cagtgtccag catctaaaaa ataggtttgg   6720 

agatggttat acaatagttg tacgaatagc agggtccaac ccggacctga agcctgtcca   6780 

ggatttcttt ggacttgcat ttcctggaag tgttctaaaa gagaaacacc ggaacatgct   6840 

acaataccag cttccatctt cattatcttc tctggccagg atattcagca tcctctccca   6900 

gagcaaaaag cgactccaca tagaagacta ctctgtttct cagacaacac ttgaccaagt   6960 

atttgtgaac tttgccaagg accaaagtga tgatgaccac ttaaaagacc tctcattaca   7020 

caaaaaccag acagtagtgg acgttgcagt tctcacatct tttctacagg atgagaaagt   7080 

gaaagaaagc tatgtatgaa gaatcctgtt catacggggt ggctgaaagt aaagaggaac   7140 

tagactttcc tttgcaccat gtgaagtgtt gtggagaaaa gagccagaag ttgatgtggg   7200 

aagaagtaaa ctggatactg tactgatact attcaatgca atgcaattca atgcaatgaa   7260 

aacaaaattc cattacaggg gcagtgcctt tgtagcctat gtcttgtatg gctctcaagt   7320 

gaaagacttg aatttagttt tttacctata cctatgtgaa actctattat ggaacccaat   7380 

ggacatatgg gtttgaactc acactttttt tttttttttt gttcctgtgt attctcattg   7440 

gggttgcaac aataattcat caagtaatca tggccagcga ttattgatca aaatcaaaag   7500 

gtaatgcaca tcctcattca ctaagccatg ccatgcccag gagactggtt tcccggtgac   7560 

acatccattg ctggcaatga gtgtgccaga gttattagtg ccaagttttt cagaaagttt   7620 

gaagcaccat ggtgtgtcat gctcactttt gtgaaagctg ctctgctcag agtctatcaa   7680 

cattgaatat cagttgacag aatggtgcca tgcgtggcta acatcctgct ttgattccct   7740 

ctgataagct gttctggtgg cagtaacatg caacaaaaat gtgggtgtct ccaggcacgg   7800 

gaaacttggt tccattgtta tattgtccta tgcttcgagc catgggtcta cagggtcatc   7860 

cttatgagac tcttaaatat acttagatcc tggtaagagg caaagaatca acagccaaac   7920 

tgctggggct gcaagctgct gaagccaggg catgggatta aagagattgt gcgttcaaac   7980 

ctagggaagc ctgtgcccat ttgtcctgac tgtctgctaa catggtacac tgcatctcaa   8040 

gatgtttatc tgacacaagt gtattatttc tggctttttg aattaatcta gaaaatgaaa   8100 

agatggagtt gtattttgac aaaaatgttt gtacttttta atgttatttg gaattttaag   8160 

ttctatcagt gacttctgaa tccttagaat ggcctctttg tagaaccctg tggtatagag   8220 

gagtatggcc actgccccac tatttttatt ttcttatgta agtttgcata tcagtcatga   8280 

ctagtgccta gaaagcaatg tgatggtcag gatctcatga cattatattt gagtttcttt   8340 

cagatcattt aggatactct taatctcact tcatcaatca aatatttttt gagtgtatgc   8400 

tgtagctgaa agagtatgta cgtacgtata agactagaga gatattaagt ctcagtacac   8460 

ttcctgtgcc atgttattca gctcactggt ttacaaatat aggttgtctt gtggttgtag   8520 

gagcccactg taacaatact gggcagcctt tttttttttt tttttaattg caacaatgca   8580 

aaagccaaga aagtataagg gtcacaagtc taaacaatga attcttcaac agggaaaaca   8640 

gctagcttga aaacttgctg aaaaacacaa cttgtgttta tggcatttag taccttcaaa   8700 

taattggctt tgcagatatt ggatacccca ttaaatctga cagtctcaaa tttttcatct   8760 

cttcaatcac tagtcaagaa aaatataaaa acaacaaata cttccatatg gagcattttt   8820 

cagagttttc taacccagtc ttatttttct agtcagtaaa catttgtaaa aatactgttt   8880 

cactaatact tactgttaac tgtcttgaga gaaaagaaaa atatgagaga actattgttt   8940 

ggggaagttc aagtgatctt tcaatatcat tactaacttc ttccactttt tccagaattt   9000 

gaatattaac gctaaaggtg taagacttca gatttcaaat taatctttct atatttttta   9060 

aatttacaga atattatata acccactgct gaaaaagaaa aaaatgattg ttttagaagt   9120 

taaagtcaat attgatttta aatataagta atgaaggcat atttccaata actagtgata   9180 

tggcatcgtt gcattttaca gtatcttcaa aaatacagaa tttatagaat aatttctcct   9240 

catttaatat ttttcaaaat caaagttatg gtttcctcat tttactaaaa tcgtattcta   9300 

attcttcatt atagtaaatc tatgagcaac tccttacttc ggttcctctg atttcaaggc   9360 

catattttaa aaaatcaaaa ggcactgtga actattttga agaaaacaca acattttaat   9420 

acagattgaa aggacctctt ctgaagctag aaacaatcta tagttataca tcttcattaa   9480 

tactgtgtta ccttttaaaa tagtaatttt ttacattttc ctgtgtaaac ctaattgtgg   9540 

tagaaatttt taccaactct atactcaatc aagcaaaatt tctgtatatt ccctgtggaa   9600 

tgtacctatg tgagtttcag aaattctcaa aatacgtgtt caaaaatttc tgcttttgca   9660 

tctttgggac acctcagaaa acttattaac aactgtgaat atgagaaata cagaagaaaa   9720 

taataagccc tctatacata aatgcccagc acaattcatt gttaaaaaac aaccaaacct   9780 

cacactactg tatttcatta tctgtactga aagcaaatgc tttgtgacta ttaaatgttg   9840 

cacatcattc attcactgta tagtaatcat tgactaaagc catttgtctg tgttttcttc   9900 

ttgtggttgt atatatcagg taaaatattt tccaaagagc catgtgtcat gtaatactga   9960 

accactttga tattgagaca ttaatttgta cccttgttat tatctactag taataatgta  10020 

atactgtaga aatattgctc taattctttt caaaattgtt gcatccccct tagaatgttt  10080 

ctatttccat aaggatttag gtatgctatt atcccttctt ataccctaag atgaagctgt  10140 

ttttgtgctc tttgttcatc attggccctc attccaagca ctttacgctg tctgtaatgg  10200 

gatctatttt tgcactggaa tatctgagaa ttgcaaaact agacaaaagt ttcacaacag  10260 

atttctaagt taaatcattt tcattaaaag gaaaaaagaa aaaaaatttt gtatgtcaat  10320 

aactttatat gaagtattaa aatgcatatt tctatgttgt aatataatga gtcacaaaat  10380 

aaagctgtga cagttctgtt ggtctacaga aa                                10412 

 
           
             8  
             6786  
             DNA  
             Human  
           
            8 

atggcttgtt ggcctcagct gaggttgctg ctgtggaaga acctcacttt cagaagaaga     60 

caaacatgtc agctgctgct ggaagtggcc tggcctctat ttatcttcct gatcctgatc    120 

tctgttcggc tgagctaccc accctatgaa caacatgaat gccattttcc aaataaagcc    180 

atgccctctg caggaacact tccttgggtt caggggatta tctgtaatgc caacaacccc    240 

tgtttccgtt acccgactcc tggggaggct cccggagttg ttggaaactt taacaaatcc    300 

attgtggctc gcctgttctc agatgctcgg aggcttcttt tatacagcca gaaagacacc    360 

agcatgaagg acatgcgcaa agttctgaga acattacagc agatcaagaa atccagctca    420 

aacttgaagc ttcaagattt cctggtggac aatgaaacct tctctgggtt cctgtatcac    480 

aacctctctc tcccaaagtc tactgtggac aagatgctga gggctgatgt cattctccac    540 

aaggtatttt tgcaaggcta ccagttacat ttgacaagtc tgtgcaatgg atcaaaatca    600 

gaagagatga ttcaacttgg tgaccaagaa gtttctgagc tttgtggcct accaagggag    660 

aaactggctg cagcagagcg agtacttcgt tccaacatgg acatcctgaa gccaatcctg    720 

agaacactaa actctacatc tcccttcccg agcaaggagc tggctgaagc cacaaaaaca    780 

ttgctgcata gtcttgggac tctggcccag gagctgttca gcatgagaag ctggagtgac    840 

atgcgacagg aggtgatgtt tctgaccaat gtgaacagct ccagctcctc cacccaaatc    900 

taccaggctg tgtctcgtat tgtctgcggg catcccgagg gaggggggct gaagatcaag    960 

tctctcaact ggtatgagga caacaactac aaagccctct ttggaggcaa tggcactgag   1020 

gaagatgctg aaaccttcta tgacaactct acaactcctt actgcaatga tttgatgaag   1080 

aatttggagt ctagtcctct ttcccgcatt atctggaaag ctctgaagcc gctgctcgtt   1140 

gggaagatcc tgtatacacc tgacactcca gccacaaggc aggtcatggc tgaggtgaac   1200 

aagaccttcc aggaactggc tgtgttccat gatctggaag gcatgtggga ggaactcagc   1260 

cccaagatct ggaccttcat ggagaacagc caagaaatgg accttgtccg gatgctgttg   1320 

gacagcaggg acaatgacca cttttgggaa cagcagttgg atggcttaga ttggacagcc   1380 

caagacatcg tggcgttttt ggccaagcac ccagaggatg tccagtccag taatggttct   1440 

gtgtacacct ggagagaagc tttcaacgag actaaccagg caatccggac catatctcgc   1500 

ttcatggagt gtgtcaacct gaacaagcta gaacccatag caacagaagt ctggctcatc   1560 

aacaagtcca tggagctgct ggatgagagg aagttctggg ctggtattgt gttcactgga   1620 

attactccag gcagcattga gctgccccat catgtcaagt acaagatccg aatggacatt   1680 

gacaatgtgg agaggacaaa taaaatcaag gatgggtact gggaccctgg tcctcgagct   1740 

gacccctttg aggacatgcg gtacgtctgg gggggcttcg cctacttgca ggatgtggtg   1800 

gagcaggcaa tcatcagggt gctgacgggc accgagaaga aaactggtgt ctatatgcaa   1860 

cagatgccct atccctgtta cgttgatgac atctttctgc gggtgatgag ccggtcaatg   1920 

cccctcttca tgacgctggc ctggatttac tcagtggctg tgatcatcaa gggcatcgtg   1980 

tatgagaagg aggcacggct gaaagagacc atgcggatca tgggcctgga caacagcatc   2040 

ctctggttta gctggttcat tagtagcctc attcctcttc ttgtgagcgc tggcctgcta   2100 

gtggtcatcc tgaagttagg aaacctgctg ccctacagtg atcccagcgt ggtgtttgtc   2160 

ttcctgtccg tgtttgctgt ggtgacaatc ctgcagtgct tcctgattag cacactcttc   2220 

tccagagcca acctggcagc agcctgtggg ggcatcatct acttcacgct gtacctgccc   2280 

tacgtcctgt gtgtggcatg gcaggactac gtgggcttca cactcaagat cttcgctagc   2340 

ctgctgtctc ctgtggcttt tgggtttggc tgtgagtact ttgccctttt tgaggagcag   2400 

ggcattggag tgcagtggga caacctgttt gagagtcctg tggaggaaga tggcttcaat   2460 

ctcaccactt cggtctccat gatgctgttt gacaccttcc tctatggggt gatgacctgg   2520 

tacattgagg ctgtctttcc aggccagtac ggaattccca ggccctggta ttttccttgc   2580 

accaagtcct actggtttgg cgaggaaagt gatgagaaga gccaccctgg ttccaaccag   2640 

aagagaatat cagaaatctg catggaggag gaacccaccc acttgaagct gggcgtgtcc   2700 

attcagaacc tggtaaaagt ctaccgagat gggatgaagg tggctgtcga tggcctggca   2760 

ctgaattttt atgagggcca gatcacctcc ttcctgggcc acaatggagc ggggaagacg   2820 

accaccatgt caatcctgac cgggttgttc cccccgacct cgggcaccgc ctacatcctg   2880 

ggaaaagaca ttcgctctga gatgagcacc atccggcaga acctgggggt ctgtccccag   2940 

cataacgtgc tgtttgacat gctgactgtc gaagaacaca tctggttcta tgcccgcttg   3000 

aaagggctct ctgagaagca cgtgaaggcg gagatggagc agatggccct ggatgttggt   3060 

ttgccatcaa gcaagctgaa aagcaaaaca agccagctgt caggtggaat gcagagaaag   3120 

ctatctgtgg ccttggcctt tgtcggggga tctaaggttg tcattctgga tgaacccaca   3180 

gctggtgtgg acccttactc ccgcagggga atatgggagc tgctgctgaa ataccgacaa   3240 

ggccgcacca ttattctctc tacacaccac atggatgaag cggacgtcct gggggacagg   3300 

attgccatca tctcccatgg gaagctgtgc tgtgtgggct cctccctgtt tctgaagaac   3360 

cagctgggaa caggctacta cctgaccttg gtcaagaaag atgtggaatc ctccctcagt   3420 

tcctgcagaa acagtagtag cactgtgtca tacctgaaaa aggaggacag tgtttctcag   3480 

agcagttctg atgctggcct gggcagcgac catgagagtg acacgctgac catcgatgtc   3540 

tctgctatct ccaacctcat caggaagcat gtgtctgaag cccggctggt ggaagacata   3600 

gggcatgagc tgacctatgt gctgccatat gaagctgcta aggagggagc ctttgtggaa   3660 

ctctttcatg agattgatga ccggctctca gacctgggca tttctagtta tggcatctca   3720 

gagacgaccc tggaagaaat attcctcaag gtggccgaag agagtggggt ggatgctgag   3780 

acctcagatg gtaccttgcc agcaagacga aacaggcggg ccttcgggga caagcagagc   3840 

tgtcttcgcc cgttcactga agatgatgct gctgatccaa atgattctga catagaccca   3900 

gaatccagag agacagactt gctcagtggg atggatggca aagggtccta ccaggtgaaa   3960 

ggctggaaac ttacacagca acagtttgtg gcccttttgt ggaagagact gctaattgcc   4020 

agacggagtc ggaaaggatt ttttgctcag attgtcttgc cagctgtgtt tgtctgcatt   4080 

gcccttgtgt tcagcctgat cgtgccaccc tttggcaagt accccagcct ggaacttcag   4140 

ccctggatgt acaacgaaca gtacacattt gtcagcaatg atgctcctga ggacacggga   4200 

accctggaac tcttaaacgc cctcaccaaa gaccctggct tcgggacccg ctgtatggaa   4260 

ggaaacccaa tcccagacac gccctgccag gcaggggagg aagagtggac cactgcccca   4320 

gttccccaga ccatcatgga cctcttccag aatgggaact ggacaatgca gaacccttca   4380 

cctgcatgcc agtgtagcag cgacaaaatc aagaagatgc tgcctgtgtg tcccccaggg   4440 

gcaggggggc tgcctcctcc acaaagaaaa caaaacactg cagatatcct tcaggacctg   4500 

acaggaagaa acatttcgga ttatctggtg aagacgtatg tgcagatcat agccaaaagc   4560 

ttaaagaaca agatctgggt gaatgagttt aggtatggcg gcttttccct gggtgtcagt   4620 

aatactcaag cacttcctcc gagtcaagaa gttaatgatg ccatcaaaca aatgaagaaa   4680 

cacctaaagc tggccaagga cagttctgca gatcgatttc tcaacagctt gggaagattt   4740 

atgacaggac tggacaccaa aaataatgtc aaggtgtggt tcaataacaa gggctggcat   4800 

gcaatcagct ctttcctgaa tgtcatcaac aatgccattc tccgggccaa cctgcaaaag   4860 

ggagagaacc ctagccatta tggaattact gctttcaatc atcccctgaa tctcaccaag   4920 

cagcagctct cagaggtggc tctgatgacc acatcagtgg atgtccttgt gtccatctgt   4980 

gtcatctttg caatgtcctt cgtcccagcc agctttgtcg tattcctgat ccaggagcgg   5040 

gtcagcaaag caaaacacct gcagttcatc agtggagtga agcctgtcat ctactggctc   5100 

tctaattttg tctgggatat gtgcaattac gttgtccctg ccacactggt cattatcatc   5160 

ttcatctgct tccagcagaa gtcctatgtg tcctccacca atctgcctgt gctagccctt   5220 

ctacttttgc tgtatgggtg gtcaatcaca cctctcatgt acccagcctc ctttgtgttc   5280 

aagatcccca gcacagccta tgtggtgctc accagcgtga acctcttcat tggcattaat   5340 

ggcagcgtgg ccacctttgt gctggagctg ttcaccgaca ataagctgaa taatatcaat   5400 

gatatcctga agtccgtgtt cttgatcttc ccacattttt gcctgggacg agggctcatc   5460 

gacatggtga aaaaccaggc aatggctgat gccctggaaa ggtttgggga gaatcgcttt   5520 

gtgtcaccat tatcttggga cttggtggga cgaaacctct tcgccatggc cgtggaaggg   5580 

gtggtgttct tcctcattac tgttctgatc cagtacagat tcttcatcag gcccagacct   5640 

gtaaatgcaa agctatctcc tctgaatgat gaagatgaag atgtgaggcg ggaaagacag   5700 

agaattcttg atggtggagg ccagaatgac atcttagaaa tcaaggagtt gacgaagata   5760 

tatagaagga agcggaagcc tgctgttgac aggatttgcg tgggcattcc tcctggtgag   5820 

tgctttgggc tcctgggagt taatggggct ggaaaatcat caactttcaa gatgttaaca   5880 

ggagatacca ctgttaccag aggagatgct ttccttaaca aaaatagtat cttatcaaac   5940 

atccatgaag tacatcagaa catgggctac tgccctcagt ttgatgccat cacagagctg   6000 

ttgactggga gagaacacgt ggagttcttt gcccttttga gaggagtccc agagaaagaa   6060 

gttggcaagg ttggtgagtg ggcgattcgg aaactgggcc tcgtgaagta tggagaaaaa   6120 

tatgctggta actatagtgg aggcaacaaa cgcaagctct ctacagccat ggctttgatc   6180 

ggcgggcctc ctgtggtgtt tctggatgaa cccaccacag gcatggatcc caaagcccgg   6240 

cggttcttgt ggaattgtgc cctaagtgtt gtcaaggagg ggagatcagt agtgcttaca   6300 

tctcatagta tggaagagtg tgaagctctt tgcactagga tggcaatcat ggtcaatgga   6360 

aggttcaggt gccttggcag tgtccagcat ctaaaaaata ggtttggaga tggttataca   6420 

atagttgtac gaatagcagg gtccaacccg gacctgaagc ctgtccagga tttctttgga   6480 

cttgcatttc ctggaagtgt tctaaaagag aaacaccgga acatgctaca ataccagctt   6540 

ccatcttcat tatcttctct ggccaggata ttcagcatcc tctcccagag caaaaagcga   6600 

ctccacatag aagactactc tgtttctcag acaacacttg accaagtatt tgtgaacttt   6660 

gccaaggacc aaagtgatga tgaccactta aaagacctct cattacacaa aaaccagaca   6720 

gtagtggacg ttgcagttct cacatctttt ctacaggatg agaaagtgaa agaaagctat   6780 

gtatga                                                              6786 

 
           
             9  
             2261  
             PRT  
             Human  
           
            9 

Met Ala Cys Trp Pro Gln Leu Arg Leu Leu Leu Trp Lys Asn Leu Thr 
1               5                   10                  15 

Phe Arg Arg Arg Gln Thr Cys Gln Leu Leu Leu Glu Val Ala Trp Pro 
            20                  25                  30 

Leu Phe Ile Phe Leu Ile Leu Ile Ser Val Arg Leu Ser Tyr Pro Pro 
        35                  40                  45 

Tyr Glu Gln His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala 
    50                  55                  60 

Gly Thr Leu Pro Trp Val Gln Gly Ile Ile Cys Asn Ala Asn Asn Pro 
65                  70                  75                  80 

Cys Phe Arg Tyr Pro Thr Pro Gly Glu Ala Pro Gly Val Val Gly Asn 
                85                  90                  95 

Phe Asn Lys Ser Ile Val Ala Arg Leu Phe Ser Asp Ala Arg Arg Leu 
            100                 105                 110 

Leu Leu Tyr Ser Gln Lys Asp Thr Ser Met Lys Asp Met Arg Lys Val 
        115                 120                 125 

Leu Arg Thr Leu Gln Gln Ile Lys Lys Ser Ser Ser Asn Leu Lys Leu 
    130                 135                 140 

Gln Asp Phe Leu Val Asp Asn Glu Thr Phe Ser Gly Phe Leu Tyr His 
145                 150                 155                 160 

Asn Leu Ser Leu Pro Lys Ser Thr Val Asp Lys Met Leu Arg Ala Asp 
                165                 170                 175 

Val Ile Leu His Lys Val Phe Leu Gln Gly Tyr Gln Leu His Leu Thr 
            180                 185                 190 

Ser Leu Cys Asn Gly Ser Lys Ser Glu Glu Met Ile Gln Leu Gly Asp 
        195                 200                 205 

Gln Glu Val Ser Glu Leu Cys Gly Leu Pro Arg Glu Lys Leu Ala Ala 
    210                 215                 220 

Ala Glu Arg Val Leu Arg Ser Asn Met Asp Ile Leu Lys Pro Ile Leu 
225                 230                 235                 240 

Arg Thr Leu Asn Ser Thr Ser Pro Phe Pro Ser Lys Glu Leu Ala Glu 
                245                 250                 255 

Ala Thr Lys Thr Leu Leu His Ser Leu Gly Thr Leu Ala Gln Glu Leu 
            260                 265                 270 

Phe Ser Met Arg Ser Trp Ser Asp Met Arg Gln Glu Val Met Phe Leu 
        275                 280                 285 

Thr Asn Val Asn Ser Ser Ser Ser Ser Thr Gln Ile Tyr Gln Ala Val 
    290                 295                 300 

Ser Arg Ile Val Cys Gly His Pro Glu Gly Gly Gly Leu Lys Ile Lys 
305                 310                 315                 320 

Ser Leu Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Leu Phe Gly Gly 
                325                 330                 335 

Asn Gly Thr Glu Glu Asp Ala Glu Thr Phe Tyr Asp Asn Ser Thr Thr 
            340                 345                 350 

Pro Tyr Cys Asn Asp Leu Met Lys Asn Leu Glu Ser Ser Pro Leu Ser 
        355                 360                 365 

Arg Ile Ile Trp Lys Ala Leu Lys Pro Leu Leu Val Gly Lys Ile Leu 
    370                 375                 380 

Tyr Thr Pro Asp Thr Pro Ala Thr Arg Gln Val Met Ala Glu Val Asn 
385                 390                 395                 400 

Lys Thr Phe Gln Glu Leu Ala Val Phe His Asp Leu Glu Gly Met Trp 
                405                 410                 415 

Glu Glu Leu Ser Pro Lys Ile Trp Thr Phe Met Glu Asn Ser Gln Glu 
            420                 425                 430 

Met Asp Leu Val Arg Met Leu Leu Asp Ser Arg Asp Asn Asp His Phe 
        435                 440                 445 

Trp Glu Gln Gln Leu Asp Gly Leu Asp Trp Thr Ala Gln Asp Ile Val 
    450                 455                 460 

Ala Phe Leu Ala Lys His Pro Glu Asp Val Gln Ser Ser Asn Gly Ser 
465                 470                 475                 480 

Val Tyr Thr Trp Arg Glu Ala Phe Asn Glu Thr Asn Gln Ala Ile Arg 
                485                 490                 495 

Thr Ile Ser Arg Phe Met Glu Cys Val Asn Leu Asn Lys Leu Glu Pro 
            500                 505                 510 

Ile Ala Thr Glu Val Trp Leu Ile Asn Lys Ser Met Glu Leu Leu Asp 
        515                 520                 525 

Glu Arg Lys Phe Trp Ala Gly Ile Val Phe Thr Gly Ile Thr Pro Gly 
    530                 535                 540 

Ser Ile Glu Leu Pro His His Val Lys Tyr Lys Ile Arg Met Asp Ile 
545                 550                 555                 560 

Asp Asn Val Glu Arg Thr Asn Lys Ile Lys Asp Gly Tyr Trp Asp Pro 
                565                 570                 575 

Gly Pro Arg Ala Asp Pro Phe Glu Asp Met Arg Tyr Val Trp Gly Gly 
            580                 585                 590 

Phe Ala Tyr Leu Gln Asp Val Val Glu Gln Ala Ile Ile Arg Val Leu 
        595                 600                 605 

Thr Gly Thr Glu Lys Lys Thr Gly Val Tyr Met Gln Gln Met Pro Tyr 
    610                 615                 620 

Pro Cys Tyr Val Asp Asp Ile Phe Leu Arg Val Met Ser Arg Ser Met 
625                 630                 635                 640 

Pro Leu Phe Met Thr Leu Ala Trp Ile Tyr Ser Val Ala Val Ile Ile 
                645                 650                 655 

Lys Gly Ile Val Tyr Glu Lys Glu Ala Arg Leu Lys Glu Thr Met Arg 
            660                 665                 670 

Ile Met Gly Leu Asp Asn Ser Ile Leu Trp Phe Ser Trp Phe Ile Ser 
        675                 680                 685 

Ser Leu Ile Pro Leu Leu Val Ser Ala Gly Leu Leu Val Val Ile Leu 
    690                 695                 700 

Lys Leu Gly Asn Leu Leu Pro Tyr Ser Asp Pro Ser Val Val Phe Val 
705                 710                 715                 720 

Phe Leu Ser Val Phe Ala Val Val Thr Ile Leu Gln Cys Phe Leu Ile 
                725                 730                 735 

Ser Thr Leu Phe Ser Arg Ala Asn Leu Ala Ala Ala Cys Gly Gly Ile 
            740                 745                 750 

Ile Tyr Phe Thr Leu Tyr Leu Pro Tyr Val Leu Cys Val Ala Trp Gln 
        755                 760                 765 

Asp Tyr Val Gly Phe Thr Leu Lys Ile Phe Ala Ser Leu Leu Ser Pro 
    770                 775                 780 

Val Ala Phe Gly Phe Gly Cys Glu Tyr Phe Ala Leu Phe Glu Glu Gln 
785                 790                 795                 800 

Gly Ile Gly Val Gln Trp Asp Asn Leu Phe Glu Ser Pro Val Glu Glu 
                805                 810                 815 

Asp Gly Phe Asn Leu Thr Thr Ser Val Ser Met Met Leu Phe Asp Thr 
            820                 825                 830 

Phe Leu Tyr Gly Val Met Thr Trp Tyr Ile Glu Ala Val Phe Pro Gly 
        835                 840                 845 

Gln Tyr Gly Ile Pro Arg Pro Trp Tyr Phe Pro Cys Thr Lys Ser Tyr 
    850                 855                 860 

Trp Phe Gly Glu Glu Ser Asp Glu Lys Ser His Pro Gly Ser Asn Gln 
865                 870                 875                 880 

Lys Arg Ile Ser Glu Ile Cys Met Glu Glu Glu Pro Thr His Leu Lys 
                885                 890                 895 

Leu Gly Val Ser Ile Gln Asn Leu Val Lys Val Tyr Arg Asp Gly Met 
            900                 905                 910 

Lys Val Ala Val Asp Gly Leu Ala Leu Asn Phe Tyr Glu Gly Gln Ile 
        915                 920                 925 

Thr Ser Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Met Ser 
    930                 935                 940 

Ile Leu Thr Gly Leu Phe Pro Pro Thr Ser Gly Thr Ala Tyr Ile Leu 
945                 950                 955                 960 

Gly Lys Asp Ile Arg Ser Glu Met Ser Thr Ile Arg Gln Asn Leu Gly 
                965                 970                 975 

Val Cys Pro Gln His Asn Val Leu Phe Asp Met Leu Thr Val Glu Glu 
            980                 985                 990 

His Ile Trp Phe Tyr Ala Arg Leu  Lys Gly Leu Ser Glu  Lys His Val 
        995                 1000                 1005 

Lys Ala  Glu Met Glu Gln Met  Ala Leu Asp Val Gly  Leu Pro Ser 
    1010                 1015                 1020 

Ser Lys  Leu Lys Ser Lys Thr  Ser Gln Leu Ser Gly  Gly Met Gln 
    1025                 1030                 1035 

Arg Lys  Leu Ser Val Ala Leu  Ala Phe Val Gly Gly  Ser Lys Val 
    1040                 1045                 1050 

Val Ile  Leu Asp Glu Pro Thr  Ala Gly Val Asp Pro  Tyr Ser Arg 
    1055                 1060                 1065 

Arg Gly  Ile Trp Glu Leu Leu  Leu Lys Tyr Arg Gln  Gly Arg Thr 
    1070                 1075                 1080 

Ile Ile  Leu Ser Thr His His  Met Asp Glu Ala Asp  Val Leu Gly 
    1085                 1090                 1095 

Asp Arg  Ile Ala Ile Ile Ser  His Gly Lys Leu Cys  Cys Val Gly 
    1100                 1105                 1110 

Ser Ser  Leu Phe Leu Lys Asn  Gln Leu Gly Thr Gly  Tyr Tyr Leu 
    1115                 1120                 1125 

Thr Leu  Val Lys Lys Asp Val  Glu Ser Ser Leu Ser  Ser Cys Arg 
    1130                 1135                 1140 

Asn Ser  Ser Ser Thr Val Ser  Tyr Leu Lys Lys Glu  Asp Ser Val 
    1145                 1150                 1155 

Ser Gln  Ser Ser Ser Asp Ala  Gly Leu Gly Ser Asp  His Glu Ser 
    1160                 1165                 1170 

Asp Thr  Leu Thr Ile Asp Val  Ser Ala Ile Ser Asn  Leu Ile Arg 
    1175                 1180                 1185 

Lys His  Val Ser Glu Ala Arg  Leu Val Glu Asp Ile  Gly His Glu 
    1190                 1195                 1200 

Leu Thr  Tyr Val Leu Pro Tyr  Glu Ala Ala Lys Glu  Gly Ala Phe 
    1205                 1210                 1215 

Val Glu  Leu Phe His Glu Ile  Asp Asp Arg Leu Ser  Asp Leu Gly 
    1220                 1225                 1230 

Ile Ser  Ser Tyr Gly Ile Ser  Glu Thr Thr Leu Glu  Glu Ile Phe 
    1235                 1240                 1245 

Leu Lys  Val Ala Glu Glu Ser  Gly Val Asp Ala Glu  Thr Ser Asp 
    1250                 1255                 1260 

Gly Thr  Leu Pro Ala Arg Arg  Asn Arg Arg Ala Phe  Gly Asp Lys 
    1265                 1270                 1275 

Gln Ser  Cys Leu Arg Pro Phe  Thr Glu Asp Asp Ala  Ala Asp Pro 
    1280                 1285                 1290 

Asn Asp  Ser Asp Ile Asp Pro  Glu Ser Arg Glu Thr  Asp Leu Leu 
    1295                 1300                 1305 

Ser Gly  Met Asp Gly Lys Gly  Ser Tyr Gln Val Lys  Gly Trp Lys 
    1310                 1315                 1320 

Leu Thr  Gln Gln Gln Phe Val  Ala Leu Leu Trp Lys  Arg Leu Leu 
    1325                 1330                 1335 

Ile Ala  Arg Arg Ser Arg Lys  Gly Phe Phe Ala Gln  Ile Val Leu 
    1340                 1345                 1350 

Pro Ala  Val Phe Val Cys Ile  Ala Leu Val Phe Ser  Leu Ile Val 
    1355                 1360                 1365 

Pro Pro  Phe Gly Lys Tyr Pro  Ser Leu Glu Leu Gln  Pro Trp Met 
    1370                 1375                 1380 

Tyr Asn  Glu Gln Tyr Thr Phe  Val Ser Asn Asp Ala  Pro Glu Asp 
    1385                 1390                 1395 

Thr Gly  Thr Leu Glu Leu Leu  Asn Ala Leu Thr Lys  Asp Pro Gly 
    1400                 1405                 1410 

Phe Gly  Thr Arg Cys Met Glu  Gly Asn Pro Ile Pro  Asp Thr Pro 
    1415                 1420                 1425 

Cys Gln  Ala Gly Glu Glu Glu  Trp Thr Thr Ala Pro  Val Pro Gln 
    1430                 1435                 1440 

Thr Ile  Met Asp Leu Phe Gln  Asn Gly Asn Trp Thr  Met Gln Asn 
    1445                 1450                 1455 

Pro Ser  Pro Ala Cys Gln Cys  Ser Ser Asp Lys Ile  Lys Lys Met 
    1460                 1465                 1470 

Leu Pro  Val Cys Pro Pro Gly  Ala Gly Gly Leu Pro  Pro Pro Gln 
    1475                 1480                 1485 

Arg Lys  Gln Asn Thr Ala Asp  Ile Leu Gln Asp Leu  Thr Gly Arg 
    1490                 1495                 1500 

Asn Ile  Ser Asp Tyr Leu Val  Lys Thr Tyr Val Gln  Ile Ile Ala 
    1505                 1510                 1515 

Lys Ser  Leu Lys Asn Lys Ile  Trp Val Asn Glu Phe  Arg Tyr Gly 
    1520                 1525                 1530 

Gly Phe  Ser Leu Gly Val Ser  Asn Thr Gln Ala Leu  Pro Pro Ser 
    1535                 1540                 1545 

Gln Glu  Val Asn Asp Ala Ile  Lys Gln Met Lys Lys  His Leu Lys 
    1550                 1555                 1560 

Leu Ala  Lys Asp Ser Ser Ala  Asp Arg Phe Leu Asn  Ser Leu Gly 
    1565                 1570                 1575 

Arg Phe  Met Thr Gly Leu Asp  Thr Lys Asn Asn Val  Lys Val Trp 
    1580                 1585                 1590 

Phe Asn  Asn Lys Gly Trp His  Ala Ile Ser Ser Phe  Leu Asn Val 
    1595                 1600                 1605 

Ile Asn  Asn Ala Ile Leu Arg  Ala Asn Leu Gln Lys  Gly Glu Asn 
    1610                 1615                 1620 

Pro Ser  His Tyr Gly Ile Thr  Ala Phe Asn His Pro  Leu Asn Leu 
    1625                 1630                 1635 

Thr Lys  Gln Gln Leu Ser Glu  Val Ala Leu Met Thr  Thr Ser Val 
    1640                 1645                 1650 

Asp Val  Leu Val Ser Ile Cys  Val Ile Phe Ala Met  Ser Phe Val 
    1655                 1660                 1665 

Pro Ala  Ser Phe Val Val Phe  Leu Ile Gln Glu Arg  Val Ser Lys 
    1670                 1675                 1680 

Ala Lys  His Leu Gln Phe Ile  Ser Gly Val Lys Pro  Val Ile Tyr 
    1685                 1690                 1695 

Trp Leu  Ser Asn Phe Val Trp  Asp Met Cys Asn Tyr  Val Val Pro 
    1700                 1705                 1710 

Ala Thr  Leu Val Ile Ile Ile  Phe Ile Cys Phe Gln  Gln Lys Ser 
    1715                 1720                 1725 

Tyr Val  Ser Ser Thr Asn Leu  Pro Val Leu Ala Leu  Leu Leu Leu 
    1730                 1735                 1740 

Leu Tyr  Gly Trp Ser Ile Thr  Pro Leu Met Tyr Pro  Ala Ser Phe 
    1745                 1750                 1755 

Val Phe  Lys Ile Pro Ser Thr  Ala Tyr Val Val Leu  Thr Ser Val 
    1760                 1765                 1770 

Asn Leu  Phe Ile Gly Ile Asn  Gly Ser Val Ala Thr  Phe Val Leu 
    1775                 1780                 1785 

Glu Leu  Phe Thr Asp Asn Lys  Leu Asn Asn Ile Asn  Asp Ile Leu 
    1790                 1795                 1800 

Lys Ser  Val Phe Leu Ile Phe  Pro His Phe Cys Leu  Gly Arg Gly 
    1805                 1810                 1815 

Leu Ile  Asp Met Val Lys Asn  Gln Ala Met Ala Asp  Ala Leu Glu 
    1820                 1825                 1830 

Arg Phe  Gly Glu Asn Arg Phe  Val Ser Pro Leu Ser  Trp Asp Leu 
    1835                 1840                 1845 

Val Gly  Arg Asn Leu Phe Ala  Met Ala Val Glu Gly  Val Val Phe 
    1850                 1855                 1860 

Phe Leu  Ile Thr Val Leu Ile  Gln Tyr Arg Phe Phe  Ile Arg Pro 
    1865                 1870                 1875 

Arg Pro  Val Asn Ala Lys Leu  Ser Pro Leu Asn Asp  Glu Asp Glu 
    1880                 1885                 1890 

Asp Val  Arg Arg Glu Arg Gln  Arg Ile Leu Asp Gly  Gly Gly Gln 
    1895                 1900                 1905 

Asn Asp  Ile Leu Glu Ile Lys  Glu Leu Thr Lys Ile  Tyr Arg Arg 
    1910                 1915                 1920 

Lys Arg  Lys Pro Ala Val Asp  Arg Ile Cys Val Gly  Ile Pro Pro 
    1925                 1930                 1935 

Gly Glu  Cys Phe Gly Leu Leu  Gly Val Asn Gly Ala  Gly Lys Ser 
    1940                 1945                 1950 

Ser Thr  Phe Lys Met Leu Thr  Gly Asp Thr Thr Val  Thr Arg Gly 
    1955                 1960                 1965 

Asp Ala  Phe Leu Asn Lys Asn  Ser Ile Leu Ser Asn  Ile His Glu 
    1970                 1975                 1980 

Val His  Gln Asn Met Gly Tyr  Cys Pro Gln Phe Asp  Ala Ile Thr 
    1985                 1990                 1995 

Glu Leu  Leu Thr Gly Arg Glu  His Val Glu Phe Phe  Ala Leu Leu 
    2000                 2005                 2010 

Arg Gly  Val Pro Glu Lys Glu  Val Gly Lys Val Gly  Glu Trp Ala 
    2015                 2020                 2025 

Ile Arg  Lys Leu Gly Leu Val  Lys Tyr Gly Glu Lys  Tyr Ala Gly 
    2030                 2035                 2040 

Asn Tyr  Ser Gly Gly Asn Lys  Arg Lys Leu Ser Thr  Ala Met Ala 
    2045                 2050                 2055 

Leu Ile  Gly Gly Pro Pro Val  Val Phe Leu Asp Glu  Pro Thr Thr 
    2060                 2065                 2070 

Gly Met  Asp Pro Lys Ala Arg  Arg Phe Leu Trp Asn  Cys Ala Leu 
    2075                 2080                 2085 

Ser Val  Val Lys Glu Gly Arg  Ser Val Val Leu Thr  Ser His Ser 
    2090                 2095                 2100 

Met Glu  Glu Cys Glu Ala Leu  Cys Thr Arg Met Ala  Ile Met Val 
    2105                 2110                 2115 

Asn Gly  Arg Phe Arg Cys Leu  Gly Ser Val Gln His  Leu Lys Asn 
    2120                 2125                 2130 

Arg Phe  Gly Asp Gly Tyr Thr  Ile Val Val Arg Ile  Ala Gly Ser 
    2135                 2140                 2145 

Asn Pro  Asp Leu Lys Pro Val  Gln Asp Phe Phe Gly  Leu Ala Phe 
    2150                 2155                 2160 

Pro Gly  Ser Val Leu Lys Glu  Lys His Arg Asn Met  Leu Gln Tyr 
    2165                 2170                 2175 

Gln Leu  Pro Ser Ser Leu Ser  Ser Leu Ala Arg Ile  Phe Ser Ile 
    2180                 2185                 2190 

Leu Ser  Gln Ser Lys Lys Arg  Leu His Ile Glu Asp  Tyr Ser Val 
    2195                 2200                 2205 

Ser Gln  Thr Thr Leu Asp Gln  Val Phe Val Asn Phe  Ala Lys Asp 
    2210                 2215                 2220 

Gln Ser  Asp Asp Asp His Leu  Lys Asp Leu Ser Leu  His Lys Asn 
    2225                 2230                 2235 

Gln Thr  Val Val Asp Val Ala  Val Leu Thr Ser Phe  Leu Gln Asp 
    2240                 2245                 2250 

Glu Lys  Val Lys Glu Ser Tyr  Val 
    2255                 2260 

 
           
             10  
             2261  
             PRT  
             Human  
           
            10 

Met Ala Cys Trp Pro Gln Leu Arg Leu Leu Leu Trp Lys Asn Leu Thr 
1               5                   10                  15 

Phe Arg Arg Arg Gln Thr Cys Gln Leu Leu Leu Glu Val Ala Trp Pro 
            20                  25                  30 

Leu Phe Ile Phe Leu Ile Leu Ile Ser Val Arg Leu Ser Tyr Pro Pro 
        35                  40                  45 

Tyr Glu Gln His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala 
    50                  55                  60 

Gly Thr Leu Pro Trp Val Gln Gly Ile Ile Cys Asn Ala Asn Asn Pro 
65                  70                  75                  80 

Cys Phe Arg Tyr Pro Thr Pro Gly Glu Ala Pro Gly Val Val Gly Asn 
                85                  90                  95 

Phe Asn Lys Ser Ile Val Ala Arg Leu Phe Ser Asp Ala Arg Arg Leu 
            100                 105                 110 

Leu Leu Tyr Ser Gln Lys Asp Thr Ser Met Lys Asp Met Arg Lys Val 
        115                 120                 125 

Leu Arg Thr Leu Gln Gln Ile Lys Lys Ser Ser Ser Asn Leu Lys Leu 
    130                 135                 140 

Gln Asp Phe Leu Val Asp Asn Glu Thr Phe Ser Gly Phe Leu Tyr His 
145                 150                 155                 160 

Asn Leu Ser Leu Pro Lys Ser Thr Val Asp Lys Met Leu Arg Ala Asp 
                165                 170                 175 

Val Ile Leu His Lys Val Phe Leu Gln Gly Tyr Gln Leu His Leu Thr 
            180                 185                 190 

Ser Leu Cys Asn Gly Ser Lys Ser Glu Glu Met Ile Gln Leu Gly Asp 
        195                 200                 205 

Gln Glu Val Ser Glu Leu Cys Gly Leu Pro Arg Glu Lys Leu Ala Ala 
    210                 215                 220 

Ala Glu Arg Val Leu Arg Ser Asn Met Asp Ile Leu Lys Pro Ile Leu 
225                 230                 235                 240 

Arg Thr Leu Asn Ser Thr Ser Pro Phe Pro Ser Lys Glu Leu Ala Glu 
                245                 250                 255 

Ala Thr Lys Thr Leu Leu His Ser Leu Gly Thr Leu Ala Gln Glu Leu 
            260                 265                 270 

Phe Ser Met Arg Ser Trp Ser Asp Met Arg Gln Glu Val Met Phe Leu 
        275                 280                 285 

Thr Asn Val Asn Ser Ser Ser Ser Ser Thr Gln Ile Tyr Gln Ala Val 
    290                 295                 300 

Ser Arg Ile Val Cys Gly His Pro Glu Gly Gly Gly Leu Lys Ile Lys 
305                 310                 315                 320 

Ser Leu Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Leu Phe Gly Gly 
                325                 330                 335 

Asn Gly Thr Glu Glu Asp Ala Glu Thr Phe Tyr Asp Asn Ser Thr Thr 
            340                 345                 350 

Pro Tyr Cys Asn Asp Leu Met Lys Asn Leu Glu Ser Ser Pro Leu Ser 
        355                 360                 365 

Arg Ile Ile Trp Lys Ala Leu Lys Pro Leu Leu Val Gly Lys Ile Leu 
    370                 375                 380 

Tyr Thr Pro Asp Thr Pro Ala Thr Arg Gln Val Met Ala Glu Val Asn 
385                 390                 395                 400 

Lys Thr Phe Gln Glu Leu Ala Val Phe His Asp Leu Glu Gly Met Trp 
                405                 410                 415 

Glu Glu Leu Ser Pro Lys Ile Trp Thr Phe Met Glu Asn Ser Gln Glu 
            420                 425                 430 

Met Asp Leu Val Arg Met Leu Leu Asp Ser Arg Asp Asn Asp His Phe 
        435                 440                 445 

Trp Glu Gln Gln Leu Asp Gly Leu Asp Trp Thr Ala Gln Asp Ile Val 
    450                 455                 460 

Ala Phe Leu Ala Lys His Pro Glu Asp Val Gln Ser Ser Asn Gly Ser 
465                 470                 475                 480 

Val Tyr Thr Trp Arg Glu Ala Phe Asn Glu Thr Asn Gln Ala Ile Arg 
                485                 490                 495 

Thr Ile Ser Arg Phe Met Glu Cys Val Asn Leu Asn Lys Leu Glu Pro 
            500                 505                 510 

Ile Ala Thr Glu Val Trp Leu Ile Asn Lys Ser Met Glu Leu Leu Asp 
        515                 520                 525 

Glu Arg Lys Phe Trp Ala Gly Ile Val Phe Thr Gly Ile Thr Pro Gly 
    530                 535                 540 

Ser Ile Glu Leu Pro His His Val Lys Tyr Lys Ile Arg Met Asp Ile 
545                 550                 555                 560 

Asp Asn Val Glu Arg Thr Asn Lys Ile Lys Asp Gly Tyr Trp Asp Pro 
                565                 570                 575 

Gly Pro Arg Ala Asp Pro Phe Glu Asp Met Arg Tyr Val Trp Gly Gly 
            580                 585                 590 

Phe Ala Tyr Leu Gln Asp Val Val Glu Gln Ala Ile Ile Arg Val Leu 
        595                 600                 605 

Thr Gly Thr Glu Lys Lys Thr Gly Val Tyr Met Gln Gln Met Pro Tyr 
    610                 615                 620 

Pro Cys Tyr Val Asp Asp Ile Phe Leu Arg Val Met Ser Arg Ser Met 
625                 630                 635                 640 

Pro Leu Phe Met Thr Leu Ala Trp Ile Tyr Ser Val Ala Val Ile Ile 
                645                 650                 655 

Lys Gly Ile Val Tyr Glu Lys Glu Ala Arg Leu Lys Glu Thr Met Arg 
            660                 665                 670 

Ile Met Gly Leu Asp Asn Ser Ile Leu Trp Phe Ser Trp Phe Ile Ser 
        675                 680                 685 

Ser Leu Ile Pro Leu Leu Val Ser Ala Gly Leu Leu Val Val Ile Leu 
    690                 695                 700 

Lys Leu Gly Asn Leu Leu Pro Tyr Ser Asp Pro Ser Val Val Phe Val 
705                 710                 715                 720 

Phe Leu Ser Val Phe Ala Val Val Thr Ile Leu Gln Cys Phe Leu Ile 
                725                 730                 735 

Ser Thr Leu Phe Ser Arg Ala Asn Leu Ala Ala Ala Cys Gly Gly Ile 
            740                 745                 750 

Ile Tyr Phe Thr Leu Tyr Leu Pro Tyr Val Leu Cys Val Ala Trp Gln 
        755                 760                 765 

Asp Tyr Val Gly Phe Thr Leu Lys Ile Phe Ala Ser Leu Leu Ser Pro 
    770                 775                 780 

Val Ala Phe Gly Phe Gly Cys Glu Tyr Phe Ala Leu Phe Glu Glu Gln 
785                 790                 795                 800 

Gly Ile Gly Val Gln Trp Asp Asn Leu Phe Glu Ser Pro Val Glu Glu 
                805                 810                 815 

Asp Gly Phe Asn Leu Thr Thr Ser Val Ser Met Met Leu Phe Asp Thr 
            820                 825                 830 

Phe Leu Tyr Gly Val Met Thr Trp Tyr Ile Glu Ala Val Phe Pro Gly 
        835                 840                 845 

Gln Tyr Gly Ile Pro Arg Pro Trp Tyr Phe Pro Cys Thr Lys Ser Tyr 
    850                 855                 860 

Trp Phe Gly Glu Glu Ser Asp Glu Lys Ser His Pro Gly Ser Asn Gln 
865                 870                 875                 880 

Lys Arg Ile Ser Glu Ile Cys Met Glu Glu Glu Pro Thr His Leu Lys 
                885                 890                 895 

Leu Gly Val Ser Ile Gln Asn Leu Val Lys Val Tyr Arg Asp Gly Met 
            900                 905                 910 

Lys Val Ala Val Asp Gly Leu Ala Leu Asn Phe Tyr Glu Gly Gln Ile 
        915                 920                 925 

Thr Ser Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Met Ser 
    930                 935                 940 

Ile Leu Thr Gly Leu Phe Pro Pro Thr Ser Gly Thr Ala Tyr Ile Leu 
945                 950                 955                 960 

Gly Lys Asp Ile Arg Ser Glu Met Ser Thr Ile Arg Gln Asn Leu Gly 
                965                 970                 975 

Val Cys Pro Gln His Asn Val Leu Phe Asp Met Leu Thr Val Glu Glu 
            980                 985                 990 

His Ile Trp Phe Tyr Ala Arg Leu  Lys Gly Leu Ser Glu  Lys His Val 
        995                 1000                 1005 

Lys Ala  Glu Met Glu Gln Met  Ala Leu Asp Val Gly  Leu Pro Ser 
    1010                 1015                 1020 

Ser Lys  Leu Lys Ser Lys Thr  Ser Gln Leu Ser Gly  Gly Met Gln 
    1025                 1030                 1035 

Arg Lys  Leu Ser Val Ala Leu  Ala Phe Val Gly Gly  Ser Lys Val 
    1040                 1045                 1050 

Val Ile  Leu Asp Glu Pro Thr  Ala Gly Val Asp Pro  Tyr Ser Arg 
    1055                 1060                 1065 

Arg Gly  Ile Trp Glu Leu Leu  Leu Lys Tyr Arg Gln  Gly Arg Thr 
    1070                 1075                 1080 

Ile Ile  Leu Ser Thr His His  Met Asp Glu Ala Asp  Val Leu Gly 
    1085                 1090                 1095 

Asp Arg  Ile Ala Ile Ile Ser  His Gly Lys Leu Cys  Cys Val Gly 
    1100                 1105                 1110 

Ser Ser  Leu Phe Leu Lys Asn  Gln Leu Gly Thr Gly  Tyr Tyr Leu 
    1115                 1120                 1125 

Thr Leu  Val Lys Lys Asp Val  Glu Ser Ser Leu Ser  Ser Cys Arg 
    1130                 1135                 1140 

Asn Ser  Ser Ser Thr Val Ser  Tyr Leu Lys Lys Glu  Asp Ser Val 
    1145                 1150                 1155 

Ser Gln  Ser Ser Ser Asp Ala  Gly Leu Gly Ser Asp  His Glu Ser 
    1160                 1165                 1170 

Asp Thr  Leu Thr Ile Asp Val  Ser Ala Ile Ser Asn  Leu Ile Arg 
    1175                 1180                 1185 

Lys His  Val Ser Glu Ala Arg  Leu Val Glu Asp Ile  Gly His Glu 
    1190                 1195                 1200 

Leu Thr  Tyr Val Leu Pro Tyr  Glu Ala Ala Lys Glu  Gly Ala Phe 
    1205                 1210                 1215 

Val Glu  Leu Phe His Glu Ile  Asp Asp Arg Leu Ser  Asp Leu Gly 
    1220                 1225                 1230 

Ile Ser  Ser Tyr Gly Ile Ser  Glu Thr Thr Leu Glu  Glu Ile Phe 
    1235                 1240                 1245 

Leu Lys  Val Ala Glu Glu Ser  Gly Val Asp Ala Glu  Thr Ser Asp 
    1250                 1255                 1260 

Gly Thr  Leu Pro Ala Arg Arg  Asn Arg Arg Ala Phe  Gly Asp Lys 
    1265                 1270                 1275 

Gln Ser  Cys Leu Arg Pro Phe  Thr Glu Asp Asp Ala  Ala Asp Pro 
    1280                 1285                 1290 

Asn Asp  Ser Asp Ile Asp Pro  Glu Ser Arg Glu Thr  Asp Leu Leu 
    1295                 1300                 1305 

Ser Gly  Met Asp Gly Lys Gly  Ser Tyr Gln Val Lys  Gly Trp Lys 
    1310                 1315                 1320 

Leu Thr  Gln Gln Gln Phe Val  Ala Leu Leu Trp Lys  Arg Leu Leu 
    1325                 1330                 1335 

Ile Ala  Arg Arg Ser Arg Lys  Gly Phe Phe Ala Gln  Ile Val Leu 
    1340                 1345                 1350 

Pro Ala  Val Phe Val Cys Ile  Ala Leu Val Phe Ser  Leu Ile Val 
    1355                 1360                 1365 

Pro Pro  Phe Gly Lys Tyr Pro  Ser Leu Glu Leu Gln  Pro Trp Met 
    1370                 1375                 1380 

Tyr Asn  Glu Gln Tyr Thr Phe  Val Ser Asn Asp Ala  Pro Glu Asp 
    1385                 1390                 1395 

Thr Gly  Thr Leu Glu Leu Leu  Asn Ala Leu Thr Lys  Asp Pro Gly 
    1400                 1405                 1410 

Phe Gly  Thr Arg Cys Met Glu  Gly Asn Pro Ile Pro  Asp Thr Pro 
    1415                 1420                 1425 

Cys Gln  Ala Gly Glu Glu Glu  Trp Thr Thr Ala Pro  Val Pro Gln 
    1430                 1435                 1440 

Thr Ile  Met Asp Leu Phe Gln  Asn Gly Asn Trp Thr  Met Gln Asn 
    1445                 1450                 1455 

Pro Ser  Pro Ala Cys Gln Cys  Ser Ser Asp Lys Ile  Lys Lys Met 
    1460                 1465                 1470 

Leu Pro  Val Cys Pro Pro Gly  Ala Gly Gly Leu Pro  Pro Pro Gln 
    1475                 1480                 1485 

Arg Lys  Gln Asn Thr Ala Asp  Ile Leu Gln Asp Leu  Thr Gly Arg 
    1490                 1495                 1500 

Asn Ile  Ser Asp Tyr Leu Val  Lys Thr Tyr Val Gln  Ile Ile Ala 
    1505                 1510                 1515 

Lys Ser  Leu Lys Asn Lys Ile  Trp Val Asn Glu Phe  Arg Tyr Gly 
    1520                 1525                 1530 

Gly Phe  Ser Leu Gly Val Ser  Asn Thr Gln Ala Leu  Pro Pro Ser 
    1535                 1540                 1545 

Gln Glu  Val Asn Asp Ala Ile  Lys Gln Met Lys Lys  His Leu Lys 
    1550                 1555                 1560 

Leu Ala  Lys Asp Ser Ser Ala  Asp Arg Phe Leu Asn  Ser Leu Gly 
    1565                 1570                 1575 

Arg Phe  Met Thr Gly Leu Asp  Thr Lys Asn Asn Val  Lys Val Trp 
    1580                 1585                 1590 

Phe Asn  Asn Lys Gly Trp His  Ala Ile Ser Ser Phe  Leu Asn Val 
    1595                 1600                 1605 

Ile Asn  Asn Ala Ile Leu Arg  Ala Asn Leu Gln Lys  Gly Glu Asn 
    1610                 1615                 1620 

Pro Ser  His Tyr Gly Ile Thr  Ala Phe Asn His Pro  Leu Asn Leu 
    1625                 1630                 1635 

Thr Lys  Gln Gln Leu Ser Glu  Val Ala Leu Met Thr  Thr Ser Val 
    1640                 1645                 1650 

Asp Val  Leu Val Ser Ile Cys  Val Ile Phe Ala Met  Ser Phe Val 
    1655                 1660                 1665 

Pro Ala  Ser Phe Val Val Phe  Leu Ile Gln Glu Arg  Val Ser Lys 
    1670                 1675                 1680 

Ala Lys  His Leu Gln Phe Ile  Ser Gly Val Lys Pro  Val Ile Tyr 
    1685                 1690                 1695 

Trp Leu  Ser Asn Phe Val Trp  Asp Met Cys Asn Tyr  Val Val Pro 
    1700                 1705                 1710 

Ala Thr  Leu Val Ile Ile Ile  Phe Ile Cys Phe Gln  Gln Lys Ser 
    1715                 1720                 1725 

Tyr Val  Ser Ser Thr Asn Leu  Pro Val Leu Ala Leu  Leu Leu Leu 
    1730                 1735                 1740 

Leu Tyr  Gly Trp Ser Ile Thr  Pro Leu Met Tyr Pro  Ala Ser Phe 
    1745                 1750                 1755 

Val Phe  Lys Ile Pro Ser Thr  Ala Tyr Val Val Leu  Thr Ser Val 
    1760                 1765                 1770 

Asn Leu  Phe Ile Gly Ile Asn  Gly Ser Val Ala Thr  Phe Val Leu 
    1775                 1780                 1785 

Glu Leu  Phe Thr Asp Asn Lys  Leu Asn Asn Ile Asn  Asp Ile Leu 
    1790                 1795                 1800 

Lys Ser  Val Phe Leu Ile Phe  Pro His Phe Cys Leu  Gly Arg Gly 
    1805                 1810                 1815 

Leu Ile  Asp Met Val Lys Asn  Gln Ala Met Ala Asp  Ala Leu Glu 
    1820                 1825                 1830 

Arg Phe  Gly Glu Asn Arg Phe  Val Ser Pro Leu Ser  Trp Asp Leu 
    1835                 1840                 1845 

Val Gly  Arg Asn Leu Phe Ala  Met Ala Val Glu Gly  Val Val Phe 
    1850                 1855                 1860 

Phe Leu  Ile Thr Val Leu Ile  Gln Tyr Arg Phe Phe  Ile Arg Pro 
    1865                 1870                 1875 

Arg Pro  Val Asn Ala Lys Leu  Ser Pro Leu Asn Asp  Glu Asp Glu 
    1880                 1885                 1890 

Asp Val  Arg Arg Glu Arg Gln  Arg Ile Leu Asp Gly  Gly Gly Gln 
    1895                 1900                 1905 

Asn Asp  Ile Leu Glu Ile Lys  Glu Leu Thr Lys Ile  Tyr Arg Arg 
    1910                 1915                 1920 

Lys Arg  Lys Pro Ala Val Asp  Arg Ile Cys Val Gly  Ile Pro Pro 
    1925                 1930                 1935 

Gly Glu  Cys Phe Gly Leu Leu  Gly Val Asn Gly Ala  Gly Lys Ser 
    1940                 1945                 1950 

Ser Thr  Phe Lys Met Leu Thr  Gly Asp Thr Thr Val  Thr Arg Gly 
    1955                 1960                 1965 

Asp Ala  Phe Leu Asn Lys Asn  Ser Ile Leu Ser Asn  Ile His Glu 
    1970                 1975                 1980 

Val His  Gln Asn Met Gly Tyr  Cys Pro Gln Phe Asp  Ala Ile Thr 
    1985                 1990                 1995 

Glu Leu  Leu Thr Gly Arg Glu  His Val Glu Phe Phe  Ala Leu Leu 
    2000                 2005                 2010 

Arg Gly  Val Pro Glu Lys Glu  Val Gly Lys Val Gly  Glu Trp Ala 
    2015                 2020                 2025 

Ile Arg  Lys Leu Gly Leu Val  Lys Tyr Gly Glu Lys  Tyr Ala Gly 
    2030                 2035                 2040 

Asn Tyr  Ser Gly Gly Asn Lys  Arg Lys Leu Ser Thr  Ala Met Ala 
    2045                 2050                 2055 

Leu Ile  Gly Gly Pro Pro Val  Val Phe Leu Asp Glu  Pro Thr Thr 
    2060                 2065                 2070 

Gly Met  Asp Pro Lys Ala Arg  Arg Phe Leu Trp Asn  Cys Ala Leu 
    2075                 2080                 2085 

Ser Val  Val Lys Glu Gly Arg  Ser Val Val Leu Thr  Ser His Ser 
    2090                 2095                 2100 

Met Glu  Glu Cys Glu Ala Leu  Cys Thr Arg Met Ala  Ile Met Val 
    2105                 2110                 2115 

Asn Gly  Arg Phe Arg Cys Leu  Gly Ser Val Gln His  Leu Lys Asn 
    2120                 2125                 2130 

Arg Phe  Gly Asp Gly Tyr Thr  Ile Val Val Arg Ile  Ala Gly Ser 
    2135                 2140                 2145 

Asn Pro  Asp Leu Lys Pro Val  Gln Asp Phe Phe Gly  Leu Ala Phe 
    2150                 2155                 2160 

Pro Gly  Ser Val Leu Lys Glu  Lys His Arg Asn Met  Leu Gln Tyr 
    2165                 2170                 2175 

Gln Leu  Pro Ser Ser Leu Ser  Ser Leu Ala Arg Ile  Phe Ser Ile 
    2180                 2185                 2190 

Leu Ser  Gln Ser Lys Lys Arg  Leu His Ile Glu Asp  Tyr Ser Val 
    2195                 2200                 2205 

Ser Gln  Thr Thr Leu Asp Gln  Val Phe Val Asn Phe  Ala Lys Asp 
    2210                 2215                 2220 

Gln Ser  Asp Asp Asp His Leu  Lys Asp Leu Ser Leu  His Lys Asn 
    2225                 2230                 2235 

Gln Thr  Val Val Asp Val Ala  Val Leu Thr Ser Phe  Leu Gln Asp 
    2240                 2245                 2250 

Glu Lys  Val Lys Glu Ser Tyr  Val 
    2255                 2260 

 
           
             11  
             2751  
             DNA  
             Human  
           
            11 

ggcacgaggc cggatccacc gtgcctctgc ggcctgcgtg cccggagtcc ccgcctgtgt     60 

cgtctctgtc gccgtccccg tctcctgcca ggcgcggagc cctgcgagcc gcgggtgggc    120 

cccaggcgcg cagacatggg ctgctccgcc aaagcgcgct gggctgccgg ggcgctgggc    180 

gtcgcggggc tactgtgcgc tgtgctgggc gctgtcatga tcgtgatggt gccgtcgctc    240 

atcaagcagc aggtccttaa gaacgtgcgc atcgacccca gtagcctgtc cttcaacatg    300 

tggaaggaga tccctatccc cttctatctc tccgtctact tctttgacgt catgaacccc    360 

agcgagatcc tgaagggcga gaagccgcag gtgcgggagc gcgggcccta cgtgtacagg    420 

gagttcaggc acaaaagcaa catcaccttc aacaacaacg acaccgtgtc cttcctcgag    480 

taccgcacct tccagttcca gccctccaag tcccacggct cggagagcga ctacatcgtc    540 

atgcccaaca tcctggtctt gggtgcggcg gtgatgatgg agaataagcc catgaccctg    600 

aagctcatca tgaccttggc attcaccacc ctcggcgaac gtgccttcat gaaccgcact    660 

gtgggtgaga tcatgtgggg ctacaaggac ccccttgtga atctcatcaa caagtacttt    720 

ccaggcatgt tccccttcaa ggacaagttc ggattatttg ctgagctcaa caactccgac    780 

tctgggctct tcacggtgtt cacgggggtc cagaacatca gcaggatcca cctcgtggac    840 

aagtggaacg ggctgagcaa ggttgacttc tggcattccg atcagtgcaa catgatcaat    900 

ggaacttctg ggcaaatgtg gccgcccttc atgactcctg agtcctcgct ggagttctac    960 

agcccggagg cctgccgatc catgaagcta atgtacaagg agtcaggggt gtttgaaggc   1020 

atccccacct atcgcttcgt ggctcccaaa accctgtttg ccaacgggtc catctaccca   1080 

cccaacgaag gcttctgccc gtgcctggag tctggaattc agaacgtcag cacctgcagg   1140 

ttcagtgccc ccttgtttct ctcccatcct cacttcctca acgccgaccc ggttctggca   1200 

gaagcggtga ctggcctgca ccctaaccag gaggcacact ccttgttcct ggacatccac   1260 

ccggtcacgg gaatccccat gaactgctct gtgaaactgc agctgagcct ctacatgaaa   1320 

tctgtcgcag gcattggaca aactgggaag attgagcctg tggtcctgcc gctgctctgg   1380 

tttgcagaga gcggggccat ggagggggag actcttcaca cattctacac tcagctggtg   1440 

ttgatgccca aggtgatgca ctatgcccag tacgtcctcc tggcgctggg ctgcgtcctg   1500 

ctgctggtcc ctgtcatctg ccaaatccgg agccaagtag gtgctggcca gagggcagcc   1560 

cgggctgaca gccattcgct tgcctgctgg gggaaagggg cctcagatcg gaccctctgg   1620 

ccaaccgcag cctggagccc acctccagca gcagtcctgc gtctctgccg gagtgggagc   1680 

ggtcactgct gggggctgcg cagcacgctt gcgtcttttg catgccgcgt tgccactact   1740 

ctgcctgttc tggaaggcct gggaccctcc cttggagggg gcacagggtc ctgaggacac   1800 

cgtgagccag ccaggcctgg ccgctgggcc tgaccggccc cccagcccct acaccccgct   1860 

tctcccggac tctcccagcg gacagccccc cagccccaca gcctgagcct cccagctgcc   1920 

atgtgcctgt tgcacacctg cacacacgcc ctggcacaca tacacacatg cgtgcaggct   1980 

tgtgcagaca ctcagggatg gagctgctgc tgaagggact tgtagggaga ggctcgtcaa   2040 

caagcactgt tctggaacct tctctccacg tggcccacag gcctgaccac aggggctgtg   2100 

ggtcctgcgt ccccttcctc gggtgagcct ggcctgtccc gttcagccgt tgggcccagg   2160 

cttcctcccc tccaaggtga aacactgcag tcccggtgtg gtggctcccc atgcaggacg   2220 

ggccaggctg ggagtgccgc cttcctgtgc caaattcagt ggggactcag tgcccaggcc   2280 

ctggccacga gctttggcct tggtctacct gccaggccag gcaaagcgcc tttacacagg   2340 

cctcggaaaa caatggagtg agcacaagat gccctgtgca gctgcccgag ggtctccgcc   2400 

caccccggcc ggactttgat ccccccgaag tcttcacagg cactgcatcg ggttgtctgg   2460 

cgcccttttc ctccagccta aactgacatc atcctatgga ctgagccggc cactctctgg   2520 

ccgaagtggc cgcaggctgt gcccccgagc tgcccccacc ccctcacagg gtccctcaga   2580 

ttataggtgc ccaggctgag gtgaagaggc ctgggggccc tgccttccgg gcgctcctgg   2640 

accctggggc aaacctgtga cccttttcta ctggaataga aatgagtttt atcatctttg   2700 

aaaaataatt cactcttgaa gtaataaacg tttaaaaaaa aaaaaaaaaa a            2751 

 
           
             12  
             552  
             PRT  
             Human  
           
            12 

Met Gly Cys Ser Ala Lys Ala Arg Trp Ala Ala Gly Ala Leu Gly Val 
1               5                   10                  15 

Ala Gly Leu Leu Cys Ala Val Leu Gly Ala Val Met Ile Val Met Val 
            20                  25                  30 

Pro Ser Leu Ile Lys Gln Gln Val Leu Lys Asn Val Arg Ile Asp Pro 
        35                  40                  45 

Ser Ser Leu Ser Phe Asn Met Trp Lys Glu Ile Pro Ile Pro Phe Tyr 
    50                  55                  60 

Leu Ser Val Tyr Phe Phe Asp Val Met Asn Pro Ser Glu Ile Leu Lys 
65                  70                  75                  80 

Gly Glu Lys Pro Gln Val Arg Glu Arg Gly Pro Tyr Val Tyr Arg Glu 
                85                  90                  95 

Phe Arg His Lys Ser Asn Ile Thr Phe Asn Asn Asn Asp Thr Val Ser 
            100                 105                 110 

Phe Leu Glu Tyr Arg Thr Phe Gln Phe Gln Pro Ser Lys Ser His Gly 
        115                 120                 125 

Ser Glu Ser Asp Tyr Ile Val Met Pro Asn Ile Leu Val Leu Gly Ala 
    130                 135                 140 

Ala Val Met Met Glu Asn Lys Pro Met Thr Leu Lys Leu Ile Met Thr 
145                 150                 155                 160 

Leu Ala Phe Thr Thr Leu Gly Glu Arg Ala Phe Met Asn Arg Thr Val 
                165                 170                 175 

Gly Glu Ile Met Trp Gly Tyr Lys Asp Pro Leu Val Asn Leu Ile Asn 
            180                 185                 190 

Lys Tyr Phe Pro Gly Met Phe Pro Phe Lys Asp Lys Phe Gly Leu Phe 
        195                 200                 205 

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

Val Gln Asn Ile Ser Arg Ile His Leu Val Asp Lys Trp Asn Gly Leu 
225                 230                 235                 240 

Ser Lys Val Asp Phe Trp His Ser Asp Gln Cys Asn Met Ile Asn Gly 
                245                 250                 255 

Thr Ser Gly Gln Met Trp Pro Pro Phe Met Thr Pro Glu Ser Ser Leu 
            260                 265                 270 

Glu Phe Tyr Ser Pro Glu Ala Cys Arg Ser Met Lys Leu Met Tyr Lys 
        275                 280                 285 

Glu Ser Gly Val Phe Glu Gly Ile Pro Thr Tyr Arg Phe Val Ala Pro 
    290                 295                 300 

Lys Thr Leu Phe Ala Asn Gly Ser Ile Tyr Pro Pro Asn Glu Gly Phe 
305                 310                 315                 320 

Cys Pro Cys Leu Glu Ser Gly Ile Gln Asn Val Ser Thr Cys Arg Phe 
                325                 330                 335 

Ser Ala Pro Leu Phe Leu Ser His Pro His Phe Leu Asn Ala Asp Pro 
            340                 345                 350 

Val Leu Ala Glu Ala Val Thr Gly Leu His Pro Asn Gln Glu Ala His 
        355                 360                 365 

Ser Leu Phe Leu Asp Ile His Pro Val Thr Gly Ile Pro Met Asn Cys 
    370                 375                 380 

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

Gly Gln Thr Gly Lys Ile Glu Pro Val Val Leu Pro Leu Leu Trp Phe 
                405                 410                 415 

Ala Glu Ser Gly Ala Met Glu Gly Glu Thr Leu His Thr Phe Tyr Thr 
            420                 425                 430 

Gln Leu Val Leu Met Pro Lys Val Met His Tyr Ala Gln Tyr Val Leu 
        435                 440                 445 

Leu Ala Leu Gly Cys Val Leu Leu Leu Val Pro Val Ile Cys Gln Ile 
    450                 455                 460 

Arg Ser Gln Val Gly Ala Gly Gln Arg Ala Ala Arg Ala Asp Ser His 
465                 470                 475                 480 

Ser Leu Ala Cys Trp Gly Lys Gly Ala Ser Asp Arg Thr Leu Trp Pro 
                485                 490                 495 

Thr Ala Ala Trp Ser Pro Pro Pro Ala Ala Val Leu Arg Leu Cys Arg 
            500                 505                 510 

Ser Gly Ser Gly His Cys Trp Gly Leu Arg Ser Thr Leu Ala Ser Phe 
        515                 520                 525 

Ala Cys Arg Val Ala Thr Thr Leu Pro Val Leu Glu Gly Leu Gly Pro 
    530                 535                 540 

Ser Leu Gly Gly Gly Thr Gly Ser 
545                 550