Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1997 Aug 15;100(4):847–854. doi: 10.1172/JCI119600

Depletion of reactive advanced glycation endproducts from diabetic uremic sera using a lysozyme-linked matrix.

T Mitsuhashi 1, Y M Li 1, S Fishbane 1, H Vlassara 1
PMCID: PMC508257  PMID: 9259584

Abstract

Diabetic uremic sera contain excessive amounts of reactive advanced glycation endproducts (AGEs), which accelerate the vasculopathy of diabetes and end-stage renal disease. To capture in vivo-derived toxic AGEs, high affinity AGE-binding protein lysozyme (LZ) was linked to a Sepharose 4B matrix. Initial studies showed that > 80% of 125I-AGE-BSA was retained by the LZ matrix, compared with < 10% retained by a control matrix. More than 60% of AGE-lysine was captured by the LZ matrix, and the LZ-bound fraction retained immunoreactivity and cross-linking activity, but had little intrinsic fluorescence (370/440 nm). After passage through the LZ matrix, AGE levels in diabetic sera (0.37+/-0.04 U/mg) were significantly reduced to a level (0.09+/-0.01 U/mg; n = 10; P < 0. 0001) comparable with the level of normal human serum, whereas total protein absorption was < 3%. The AGE-enriched serum fraction exhibited cross-linking activity, which was completely prevented by aminoguanidine. Among numerous LZ-bound proteins in diabetic uremic sera, three major proteins "susceptible" to AGE modification were identified: the immunoglobulin G light chain, apolipoprotein J (clusterin/SP-40,40), and the complement 3b beta chain. These findings indicate that the LZ-linked AGE affinity column may serve as an efficient method for the depletion of toxic AGEs from sera, including specific AGE-modified proteins that may be linked to altered immunity, lipoprotein metabolism, and accelerated vasculopathy in renal failure patients with or without diabetes.

Full Text

The Full Text of this article is available as a PDF (349.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ahmed M. U., Thorpe S. R., Baynes J. W. Identification of N epsilon-carboxymethyllysine as a degradation product of fructoselysine in glycated protein. J Biol Chem. 1986 Apr 15;261(11):4889–4894. [PubMed] [Google Scholar]
  2. Anderson B. F., Baker H. M., Norris G. E., Rumball S. V., Baker E. N. Apolactoferrin structure demonstrates ligand-induced conformational change in transferrins. Nature. 1990 Apr 19;344(6268):784–787. doi: 10.1038/344784a0. [DOI] [PubMed] [Google Scholar]
  3. Brownlee M., Cerami A., Vlassara H. Advanced glycosylation end products in tissue and the biochemical basis of diabetic complications. N Engl J Med. 1988 May 19;318(20):1315–1321. doi: 10.1056/NEJM198805193182007. [DOI] [PubMed] [Google Scholar]
  4. Brownlee M. Lilly Lecture 1993. Glycation and diabetic complications. Diabetes. 1994 Jun;43(6):836–841. doi: 10.2337/diab.43.6.836. [DOI] [PubMed] [Google Scholar]
  5. Brownlee M., Vlassara H., Kooney A., Ulrich P., Cerami A. Aminoguanidine prevents diabetes-induced arterial wall protein cross-linking. Science. 1986 Jun 27;232(4758):1629–1632. doi: 10.1126/science.3487117. [DOI] [PubMed] [Google Scholar]
  6. Bucala R., Makita Z., Koschinsky T., Cerami A., Vlassara H. Lipid advanced glycosylation: pathway for lipid oxidation in vivo. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6434–6438. doi: 10.1073/pnas.90.14.6434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Buttyan R., Olsson C. A., Pintar J., Chang C., Bandyk M., Ng P. Y., Sawczuk I. S. Induction of the TRPM-2 gene in cells undergoing programmed death. Mol Cell Biol. 1989 Aug;9(8):3473–3481. doi: 10.1128/mcb.9.8.3473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Choi N. H., Mazda T., Tomita M. A serum protein SP40,40 modulates the formation of membrane attack complex of complement on erythrocytes. Mol Immunol. 1989 Sep;26(9):835–840. doi: 10.1016/0161-5890(89)90139-9. [DOI] [PubMed] [Google Scholar]
  9. Collard M. W., Griswold M. D. Biosynthesis and molecular cloning of sulfated glycoprotein 2 secreted by rat Sertoli cells. Biochemistry. 1987 Jun 16;26(12):3297–3303. doi: 10.1021/bi00386a008. [DOI] [PubMed] [Google Scholar]
  10. Danik M., Chabot J. G., Mercier C., Benabid A. L., Chauvin C., Quirion R., Suh M. Human gliomas and epileptic foci express high levels of a mRNA related to rat testicular sulfated glycoprotein 2, a purported marker of cell death. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8577–8581. doi: 10.1073/pnas.88.19.8577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dolhofer R., Siess E. A., Wieland O. H. Nonenzymatic glycation of immunoglobulins leads to an impairment of immunoreactivity. Biol Chem Hoppe Seyler. 1985 Apr;366(4):361–366. doi: 10.1515/bchm3.1985.366.1.361. [DOI] [PubMed] [Google Scholar]
  12. Ghiso J., Matsubara E., Koudinov A., Choi-Miura N. H., Tomita M., Wisniewski T., Frangione B. The cerebrospinal-fluid soluble form of Alzheimer's amyloid beta is complexed to SP-40,40 (apolipoprotein J), an inhibitor of the complement membrane-attack complex. Biochem J. 1993 Jul 1;293(Pt 1):27–30. doi: 10.1042/bj2930027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hennessey P. J., Black C. T., Andrassy R. J. Nonenzymatic glycosylation of immunoglobulin G impairs complement fixation. JPEN J Parenter Enteral Nutr. 1991 Jan-Feb;15(1):60–64. doi: 10.1177/014860719101500160. [DOI] [PubMed] [Google Scholar]
  14. James R. W., Hochstrasser A. C., Borghini I., Martin B., Pometta D., Hochstrasser D. Characterization of a human high density lipoprotein-associated protein, NA1/NA2. Identity with SP-40,40, an inhibitor of complement-mediated cytolysis. Arterioscler Thromb. 1991 May-Jun;11(3):645–652. doi: 10.1161/01.atv.11.3.645. [DOI] [PubMed] [Google Scholar]
  15. Kelso G. J., Stuart W. D., Richter R. J., Furlong C. E., Jordan-Starck T. C., Harmony J. A. Apolipoprotein J is associated with paraoxonase in human plasma. Biochemistry. 1994 Jan 25;33(3):832–839. doi: 10.1021/bi00169a026. [DOI] [PubMed] [Google Scholar]
  16. Kennedy D. M., Skillen A. W., Self C. H. Glycation of monoclonal antibodies impairs their ability to bind antigen. Clin Exp Immunol. 1994 Nov;98(2):245–251. doi: 10.1111/j.1365-2249.1994.tb06133.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Koschinsky T., He C. J., Mitsuhashi T., Bucala R., Liu C., Buenting C., Heitmann K., Vlassara H. Orally absorbed reactive glycation products (glycotoxins): an environmental risk factor in diabetic nephropathy. Proc Natl Acad Sci U S A. 1997 Jun 10;94(12):6474–6479. doi: 10.1073/pnas.94.12.6474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kume S., Takeya M., Mori T., Araki N., Suzuki H., Horiuchi S., Kodama T., Miyauchi Y., Takahashi K. Immunohistochemical and ultrastructural detection of advanced glycation end products in atherosclerotic lesions of human aorta with a novel specific monoclonal antibody. Am J Pathol. 1995 Sep;147(3):654–667. [PMC free article] [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. Li Y. M., Tan A. X., Vlassara H. Antibacterial activity of lysozyme and lactoferrin is inhibited by binding of advanced glycation-modified proteins to a conserved motif. Nat Med. 1995 Oct;1(10):1057–1061. doi: 10.1038/nm1095-1057. [DOI] [PubMed] [Google Scholar]
  21. Makita Z., Bucala R., Rayfield E. J., Friedman E. A., Kaufman A. M., Korbet S. M., Barth R. H., Winston J. A., Fuh H., Manogue K. R. Reactive glycosylation endproducts in diabetic uraemia and treatment of renal failure. Lancet. 1994 Jun 18;343(8912):1519–1522. doi: 10.1016/s0140-6736(94)92935-1. [DOI] [PubMed] [Google Scholar]
  22. Makita Z., Radoff S., Rayfield E. J., Yang Z., Skolnik E., Delaney V., Friedman E. A., Cerami A., Vlassara H. Advanced glycosylation end products in patients with diabetic nephropathy. N Engl J Med. 1991 Sep 19;325(12):836–842. doi: 10.1056/NEJM199109193251202. [DOI] [PubMed] [Google Scholar]
  23. Makita Z., Vlassara H., Cerami A., Bucala R. Immunochemical detection of advanced glycosylation end products in vivo. J Biol Chem. 1992 Mar 15;267(8):5133–5138. [PubMed] [Google Scholar]
  24. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  25. Mitsuhashi T., Nakayama H., Itoh T., Kuwajima S., Aoki S., Atsumi T., Koike T. Immunochemical detection of advanced glycation end products in renal cortex from STZ-induced diabetic rat. Diabetes. 1993 Jun;42(6):826–832. doi: 10.2337/diab.42.6.826. [DOI] [PubMed] [Google Scholar]
  26. Miyata T., Oda O., Inagi R., Iida Y., Araki N., Yamada N., Horiuchi S., Taniguchi N., Maeda K., Kinoshita T. beta 2-Microglobulin modified with advanced glycation end products is a major component of hemodialysis-associated amyloidosis. J Clin Invest. 1993 Sep;92(3):1243–1252. doi: 10.1172/JCI116696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nakamura Y., Horii Y., Nishino T., Shiiki H., Sakaguchi Y., Kagoshima T., Dohi K., Makita Z., Vlassara H., Bucala R. Immunohistochemical localization of advanced glycosylation end products in coronary atheroma and cardiac tissue in diabetes mellitus. Am J Pathol. 1993 Dec;143(6):1649–1656. [PMC free article] [PubMed] [Google Scholar]
  28. Palmer D. J., Christie D. L. The primary structure of glycoprotein III from bovine adrenal medullary chromaffin granules. Sequence similarity with human serum protein-40,40 and rat Sertoli cell glycoprotein. J Biol Chem. 1990 Apr 25;265(12):6617–6623. [PubMed] [Google Scholar]
  29. Papanastasiou P., Grass L., Rodela H., Patrikarea A., Oreopoulos D., Diamandis E. P. Immunological quantification of advanced glycosylation end-products in the serum of patients on hemodialysis or CAPD. Kidney Int. 1994 Jul;46(1):216–222. doi: 10.1038/ki.1994.262. [DOI] [PubMed] [Google Scholar]
  30. Sell D. R., Monnier V. M. Structure elucidation of a senescence cross-link from human extracellular matrix. Implication of pentoses in the aging process. J Biol Chem. 1989 Dec 25;264(36):21597–21602. [PubMed] [Google Scholar]
  31. Skolnik E. Y., Yang Z., Makita Z., Radoff S., Kirstein M., Vlassara H. Human and rat mesangial cell receptors for glucose-modified proteins: potential role in kidney tissue remodelling and diabetic nephropathy. J Exp Med. 1991 Oct 1;174(4):931–939. doi: 10.1084/jem.174.4.931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tenovuo J., Lumikari M., Soukka T. Salivary lysozyme, lactoferrin and peroxidases: antibacterial effects on cariogenic bacteria and clinical applications in preventive dentistry. Proc Finn Dent Soc. 1991;87(2):197–208. [PubMed] [Google Scholar]
  33. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Vlassara H., Bucala R., Striker L. Pathogenic effects of advanced glycosylation: biochemical, biologic, and clinical implications for diabetes and aging. Lab Invest. 1994 Feb;70(2):138–151. [PubMed] [Google Scholar]
  35. Vlassara H., Fuh H., Donnelly T., Cybulsky M. Advanced glycation endproducts promote adhesion molecule (VCAM-1, ICAM-1) expression and atheroma formation in normal rabbits. Mol Med. 1995 May;1(4):447–456. [PMC free article] [PubMed] [Google Scholar]
  36. Vlassara H., Fuh H., Makita Z., Krungkrai S., Cerami A., Bucala R. Exogenous advanced glycosylation end products induce complex vascular dysfunction in normal animals: a model for diabetic and aging complications. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12043–12047. doi: 10.1073/pnas.89.24.12043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Zlokovic B. V., Martel C. L., Mackic J. B., Matsubara E., Wisniewski T., McComb J. G., Frangione B., Ghiso J. Brain uptake of circulating apolipoproteins J and E complexed to Alzheimer's amyloid beta. Biochem Biophys Res Commun. 1994 Dec 15;205(2):1431–1437. doi: 10.1006/bbrc.1994.2825. [DOI] [PubMed] [Google Scholar]
  38. de Silva H. V., Stuart W. D., Duvic C. R., Wetterau J. R., Ray M. J., Ferguson D. G., Albers H. W., Smith W. R., Harmony J. A. A 70-kDa apolipoprotein designated ApoJ is a marker for subclasses of human plasma high density lipoproteins. J Biol Chem. 1990 Aug 5;265(22):13240–13247. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES