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. 1985 Sep;82(17):5588–5592. doi: 10.1073/pnas.82.17.5588

High-affinity-receptor-mediated uptake and degradation of glucose-modified proteins: a potential mechanism for the removal of senescent macromolecules.

H Vlassara, M Brownlee, A Cerami
PMCID: PMC390596  PMID: 2994035

Abstract

Proteins that have been modified by long-term exposure to glucose accumulate advanced glycosylation end products (AGE) as a function of protein age. In these studies, we have characterized the interaction of AGE-protein with mouse peritoneal macrophages, using AGE-modified bovine serum albumin (AGE-BSA, prepared by incubation with glucose) as a probe. AGE-BSA was specifically bound to cells at 4 degrees C and was taken up and degraded at 37 degrees C; these processes were concentration dependent and saturable. Competition experiments with AGE-BSA, BSA incubated with phosphate-buffered saline rather than glucose, and yeast mannan demonstrated that macrophages specifically recognize AGE on proteins by a receptor that is completely distinct from the mannose/fucose receptor. Scatchard analysis of AGE-BSA binding data indicated that there are approximately 1.06 X 10(5) receptors per macrophage, with an affinity constant of 1.75 X 10(-11) M. Specific binding of AGE-BSA to the macrophage receptor was competitively inhibited by BSA that had been chemically coupled to a synthetic analogue of the specific AGE, 2-(2-furoyl)-4(5)-(2-furanyl)-1H-imidazole (FFI-BSA). FFI-BSA was also taken up by macrophages in a concentration-dependent, saturable manner. Prior incubation of macrophages with AGE-BSA failed to influence the subsequent uptake and degradation of added AGE-BSA. Thus, the AGE receptor does not appear to be down-regulated by exposure to AGE-proteins. Results from these studies suggest that AGE could act in vivo as a specific signal for recognition and degradation of senescent macromolecules. Incomplete removal of AGE-proteins by macrophages may ultimately give rise to some of the physiologic changes that occur with normal aging.

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Selected References

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  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Brownlee M., Vlassara H., Cerami A. Nonenzymatic glycosylation and the pathogenesis of diabetic complications. Ann Intern Med. 1984 Oct;101(4):527–537. doi: 10.7326/0003-4819-101-4-527. [DOI] [PubMed] [Google Scholar]
  3. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  4. Imber M. J., Pizzo S. V., Johnson W. J., Adams D. O. Selective diminution of the binding of mannose by murine macrophages in the late stages of activation. J Biol Chem. 1982 May 10;257(9):5129–5135. [PubMed] [Google Scholar]
  5. Koenig R. J., Cerami A. Hemoglobin A Ic and diabetes mellitus. Annu Rev Med. 1980;31:29–34. doi: 10.1146/annurev.me.31.020180.000333. [DOI] [PubMed] [Google Scholar]
  6. Monnier V. M., Kohn R. R., Cerami A. Accelerated age-related browning of human collagen in diabetes mellitus. Proc Natl Acad Sci U S A. 1984 Jan;81(2):583–587. doi: 10.1073/pnas.81.2.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Mortensen H. B., Christophersen C. Glucosylation of human haemoglobin a in red blood cells studied in vitro. Kinetics of the formation and dissociation of haemoglobin A1c. Clin Chim Acta. 1983 Nov 15;134(3):317–326. doi: 10.1016/0009-8981(83)90370-4. [DOI] [PubMed] [Google Scholar]
  8. Pongor S., Ulrich P. C., Bencsath F. A., Cerami A. Aging of proteins: isolation and identification of a fluorescent chromophore from the reaction of polypeptides with glucose. Proc Natl Acad Sci U S A. 1984 May;81(9):2684–2688. doi: 10.1073/pnas.81.9.2684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. REYNOLDS T. M. CHEMISTRY OF NONENZYMIC BROWNING. I. THE REACTION BETWEEN ALDOSES AND AMINES. Adv Food Res. 1963;12:1–52. doi: 10.1016/s0065-2628(08)60005-1. [DOI] [PubMed] [Google Scholar]
  10. Reynolds T. M. Chemistry of nonenzymic browning. II. Adv Food Res. 1965;14:167–283. doi: 10.1016/s0065-2628(08)60149-4. [DOI] [PubMed] [Google Scholar]
  11. Schnider S. L., Kohn R. R. Effects of age and diabetes mellitus on the solubility and nonenzymatic glucosylation of human skin collagen. J Clin Invest. 1981 Jun;67(6):1630–1635. doi: 10.1172/JCI110198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Schnider S. L., Kohn R. R. Glucosylation of human collagen in aging and diabetes mellitus. J Clin Invest. 1980 Nov;66(5):1179–1181. doi: 10.1172/JCI109950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Unkeless J. C. The presence of two Fc receptors on mouse macrophages: evidence from a variant cell line and differential trypsin sensitivity. J Exp Med. 1977 Apr 1;145(4):931–945. doi: 10.1084/jem.145.4.931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Vlassara H., Brownlee M., Cerami A. Accumulation of diabetic rat peripheral nerve myelin by macrophages increases with the presence of advanced glycosylation endproducts. J Exp Med. 1984 Jul 1;160(1):197–207. doi: 10.1084/jem.160.1.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Vlassara H., Brownlee M., Cerami A. Nonenzymatic glycosylation of peripheral nerve protein in diabetes mellitus. Proc Natl Acad Sci U S A. 1981 Aug;78(8):5190–5192. doi: 10.1073/pnas.78.8.5190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Vlassara H., Brownlee M., Cerami A. Recognition and uptake of human diabetic peripheral nerve myelin by macrophages. Diabetes. 1985 Jun;34(6):553–557. doi: 10.2337/diab.34.6.553. [DOI] [PubMed] [Google Scholar]

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