Claims
- 1. A receptor protein derived from rat liver cell membranes, that recognizes and binds advanced glycosylation endproducts and that possesses the following characteristics:
- A. it recognizes and binds with the ligands AGE-Rnase and AGE-Collagen I;
- B. it does not recognize and bind with the ligands FFI-BSA, formaldehyde-treated BSA, glucosamide-BSA, and acetyl LDL-BSA in a solid phase ligand blotting assay; and
- C. it has a molecular mass of about 90 kD or about 60 kD as determined by the migration of the protein on SDS-PAGE.
- 2. A receptor protein in purified form selected from the group consisting of a first protein having a molecular mass of about 90 kD and a second protein having a molecular mass of about 60 kD as determined by migration on SDS-PAGE, or a mixture of said purified first and second proteins; each of said proteins having the following characteristics:
- A. said protein is found on rat liver cell membranes;
- B. said protein is expressed on rat monocytes and rat macrophages;
- C. said protein copurifies from elutions based on an AGE ligand affinity column, an anion exchange column, and a hydroxylapatite column; and also bear the following distinction:
- D. when said protein is immobilized on nitrocellulose in a solid phase assay, the 90 kD protein does not bind to AGE-modified ligands while the 60 kD protein does.
- 3. The receptor protein of claim 1 isolated and purified by the immobilization of detergent-solubilized rat liver membrane proteins on nitrocellulose.
- 4. The receptor protein of claim 1 having the NH.sub.2 -terminal partial amino acid sequence for the said 90 kD component protein as set forth in FIG. 11 and SEQ. ID NO. 1.
- 5. The receptor protein of claim 1 having the NH.sub.2 -terminal partial amino acid sequence for the said 60 kD component protein as set forth in FIG. 12 and SEQ. ID No. 2.
- 6. A protein in purified form having a molecular mass of about 90 kD and exhibiting activity as a receptor for advanced glycosylation endproducts having the NH.sub.2 -terminal partial amino acid sequence set forth in FIG. 11 SEQ. ID. No:1.
- 7. A protein in purified form having a molecular mass of about 60 kD and exhibiting activity as a receptor for advanced glycosylation endproducts having the NH.sub.2 -terminal partial amino acid sequence set forth in FIG. 12 SEQ ID No:2.
- 8. The protein as defined by any of claims 1-5 labeled with a detectable label.
- 9. The protein of either of claims 6 or 7 labeled with a detectable label.
- 10. The receptor protein of claim 8 wherein the label is selected from enzymes, chemicals which fluoresce and radioactive elements.
- 11. The protein of claim 9 wherein the label is selected from enzymes, chemicals which fluoresce and radioactive elements.
- 12. A composition comprising:
- A. a receptor protein in purified form selected from the group consisting of a first protein having a molecular mass of about 90 kD and a second protein having a molecular mass of about 60 kD as determined by migration on SDS-PAGE, or a mixture of said purified first and second proteins; each of said proteins being characterized as derived from rat liver cell membranes, recognizing and binding to advanced glycosylation endproducts (AGE) and possessing the following characteristics:
- i. each recognizes and binds with the ligands AGE-RNase and AGE-collagen I; and
- ii. each does not recognize and bind with the ligands FFI-BSA, formaldehyde-treated BSA, glucosamide-BSA, and acetyl LDL-BSA in a solid phase ligand clotting assay; and
- B. a pharmaceutically acceptable carrier.
- 13. The composition of claim 12 containing at least one receptor protein having a molecular mass of about 90 kD and a second receptor protein having a molecular mass of about 60 kD, each of said proteins having the following characteristics:
- A. each is found on rat liver cell membranes;
- B. each is expressed on rat monocytes and rat macrophages;
- C. each copurifies from elutions based on an AGE ligand affinity column, an anion exchange column, and a hydroxylapatite column; and also bear the following distinction:
- D. when said protein is immobilized on nitrate, cellulose in a solid phase assay, the 90 kD protein does not bind to AGE-modified ligands while the 60 kD protein does.
- 14. The composition of claim 12 wherein said protein is isolated and purified by the immobilization of detergentosolubilized rat liver cell membrane proteins on nitrocellulose.
- 15. The composition of claim 12 wherein the said 90 kD protein has the NH.sub.2 -terminal partial amino acid sequence set forth in FIG. 11.
- 16. The composition of claim 12 wherein the said 60 kD protein has the NH.sub.2 -terminal partial amino acid sequence set forth in FIG. 12.
CROSS-REFERENCE TO RELATED APPLICATIONS
The present Application is a Continuation-In-Part of application Ser. No. 453,958, filed Dec. 20, 1989, now abandoned, which is in turn, a Division of application Ser. No. 091,534, filed Sep. 3, 1987, now U.S. Pat. No. 4,900,747, issued Feb. 13, 1990, which is in turn, a Continuation-In-Part of application Ser. No. 907,747, filed Sep. 12, 1986, now abandoned; all of the above preceding applications by Helen Vlassara, Michael Brownlee and Anthony Cerami, said Ser. No. 907,747, in turn, a Continuation-In-Part of application Ser. No. 798,032, filed Nov. 14, 1985, by Anthony Cerami, Peter Ulrich and Michael Brownlee, now U.S. Pat. No. 4,758,583, which is, in turn, a Continuation-In-Part of application Ser. No. 590,820, now U.S. Pat. No. 4,665,192, filed Mar. 19, 1984 by Anthony Cerami alone.
Priority under 35 U.S.C. .sctn.120 is claimed as to all of the above earlier filed Applications, and the disclosures thereof are incorporated herein by reference.
Government Interests
This invention was made with partial assistance from grant Nos. AG 8245 and DK 19655 from the National Institutes of Health. The government may have certain rights in this invention.
US Referenced Citations (1)
Number |
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Date |
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5202424 |
Vlassara et al. |
Apr 1993 |
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Yang, Z., et al. J. Exp. Med. 174:515-524 published on Aug. 23, 1991. |
Vlassara, H. et al. Proc. Natl. Acad. Sci. U.S.A. 82:5588 (1985). |
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M. Kirstein et al., Advanced protein glycosylation induces transendothelial human monocyte chemotaxis and secretion of platelet-derived growth factor: Role in vascular disease of diabetes and aging, Proc. Natl. Acad. Sci. USA, 87:9010-9014, Nov. 1990. |
Divisions (1)
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91534 |
Sep 1987 |
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Continuation in Parts (4)
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453958 |
Dec 1989 |
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907747 |
Sep 1986 |
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798032 |
Nov 1985 |
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590820 |
Mar 1984 |
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