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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

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Volume 52, Issue 1 (Jan 2014)


Site-specific AGE modifications in the extracellular matrix: a role for glyoxal in protein damage in diabetes

Paul Voziyan
  • Corresponding author
  • Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, S-3223 MCN, 1161 21st Avenue South, Nashville, TN 37232–2372, USA
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Kyle L. Brown / Sergei Chetyrkin / Billy Hudson
  • Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA
  • Department of Biochemistry, Vanderbilt University Medical Center, Nashville, TN, USA
  • Department of Pathology, Vanderbilt University Medical Center, Nashville, TN, USA
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2013-03-13 | DOI: https://doi.org/10.1515/cclm-2012-0818


Non-enzymatic modification of proteins in hyperglycemia is a major proposed mechanism of diabetic complications. Specifically, advanced glycation end products (AGEs) derived from hyperglycemia-induced reactive carbonyl species (RCS) can have pathogenic consequences when they target functionally critical protein residues. Modification of a small number of these critical residues, often undetectable by the methodologies relying on measurements of total AGE levels, can cause significant functional damage. Therefore, detection of specific sites of protein damage in diabetes is central to understanding the molecular basis of diabetic complications and for identification of biomarkers which are mechanistically linked to the disease. The current paradigm of RCS-derived protein damage places a major focus on methylglyoxal (MGO), an intermediate of cellular glycolysis. We propose that glyoxal (GO) is a major contributor to extracellular matrix (ECM) damage in diabetes. Here, we review the current knowledge and provide new data about GO-derived site-specific ECM modification in experimental diabetes.

Keywords: advanced glycation end products; collagen IV; diabetes; extracellular matrix; glyoxal; mass spectrometry; methylglyoxal; pyridoxamine


  • 1.

    Brownlee M. The pathobiology of diabetic complications: a unifying mechanism. Diabetes 2005;54:1615–25.PubMedCrossrefGoogle Scholar

  • 2.

    Lo TW, Westwood ME, McLellan AC, Selwood T, Thornalley PJ. Binding and modification of proteins by methylglyoxal under physiological conditions. A kinetic and mechanistic study with N α-acetylarginine, N α-acetylcysteine, and N α-acetyllysine, and bovine serum albumin. J Biol Chem 1994;269:32299–305.Google Scholar

  • 3.

    Rabbani N, Thornalley PJ. Methylglyoxal, glyoxalase 1 and the dicarbonyl proteome. Amino Acids 2012;42:1133–42.PubMedGoogle Scholar

  • 4.

    Yao D, Taguchi T, Matsumura T, Pestell R, Edelstein D, Giardino I, et al. High glucose increases angiopoietin-2 transcription in microvascular endothelial cells through methylglyoxal modification of mSin3A. J Biol Chem 2007;282:31038–45.Google Scholar

  • 5.

    Queisser MA, Yao D, Geisler S, Hammes HP, Lochnit G, Schleicher ED, et al. Hyperglycemia impairs proteasome function by methylglyoxal. Diabetes 2010;59:670–8.PubMedCrossrefGoogle Scholar

  • 6.

    Dobler D, Ahmed N, Song L, Eboigbodin KE, Thornalley PJ. Increased dicarbonyl metabolism in endothelial cells in hyperglycemia induces anoikis and impairs angiogenesis by RGD and GFOGER motif modification. Diabetes 2006;55:1961–9.PubMedCrossrefGoogle Scholar

  • 7.

    Wells-Knecht KJ, Zyzak DV, Litchfield JE, Thorpe SR, Baynes JW. Mechanism of autoxidative glycosylation: identification of glyoxal and arabinose as intermediates in the autoxidative modification of proteins by glucose. Biochemistry 1995;34:3702–9.CrossrefGoogle Scholar

  • 8.

    Thornalley PJ, Langborg A, Minhas HS. Formation of glyoxal, methylglyoxal and 3-deoxyglucosone in the glycation of proteins by glucose. Biochem J 1999;344:109–16.Google Scholar

  • 9.

    Chetyrkin S, Mathis M, Pedchenko V, Sanchez OA, McDonald WH, Hachey DL, et al. Glucose autoxidation induces functional damage to proteins via modification of critical arginine residues. Biochemistry 2011;50:6102–12.CrossrefPubMedGoogle Scholar

  • 10.

    Fu MX, Requena JR, Jenkins AJ, Lyons TJ, Baynes JW, Thorpe SR. The advanced glycation end product, Nε-(carboxymethyl)lysine, is a product of both lipid peroxidation and glycoxidation reactions. J Biol Chem 1996;271:9982–6.Google Scholar

  • 11.

    Lange JN, Wood KD, Knight J, Assimos DG, Holmes RP. Glyoxal formation and its role in endogenous oxalate synthesis. Adv Urol 2012;2012:819202.PubMedGoogle Scholar

  • 12.

    Hazen SL, d’Avignon A, Anderson MM, Hsu FF, Heinecke JW. Human neutrophils employ the myeloperoxidase-hydrogen peroxide-chloride system to oxidize α-amino acids to a family of reactive aldehydes. Mechanistic studies identifying labile intermediates along the reaction pathway. J Biol Chem 1998;273:4997–5005.Google Scholar

  • 13.

    Yoon KD, Yamamoto K, Ueda K, Zhou J, Sparrow JR. A novel source of methylglyoxal and glyoxal in retina: implications for age-related macular degeneration. PLoS One 2012;7:e41309.PubMedGoogle Scholar

  • 14.

    Glomb MA, Monnier VM. Mechanism of protein modification by glyoxal and glycolaldehyde, reactive intermediates of the Maillard reaction. J Biol Chem 1995;270:10017–26.Google Scholar

  • 15.

    Ferreira AE, Ponces Freire AM, Voit EO. A quantitative model of the generation of Nε-(carboxymethyl)lysine in the Maillard reaction between collagen and glucose. Biochem J 2003;376:109–21.Google Scholar

  • 16.

    Thorpe SR, Baynes JW. Maillard reaction products in tissue proteins: new products and new perspectives. Amino Acids 2003;25:275–81.PubMedCrossrefGoogle Scholar

  • 17.

    Biemel KM, Friedl DA, Lederer MO. Identification and quantification of major Maillard cross-links in human serum albumin and lens protein. Evidence for glucosepane as the dominant compound. J Biol Chem 2002;277:24907–15.Google Scholar

  • 18.

    Ahmed N, Thornalley PJ. Chromatographic assay of glycation adducts in human serum albumin glycated in vitro by derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate and intrinsic fluorescence. Biochem J 2002;364:15–24.Google Scholar

  • 19.

    Perkins BA, Rabbani N, Weston A, Ficociello LH, Adaikalakoteswari A, Niewczas M, et al. Serum levels of advanced glycation endproducts and other markers of protein damage in early diabetic nephropathy in type 1 diabetes. PLoS One 2012;7:e35655.PubMedGoogle Scholar

  • 20.

    Jabeen R, Saleemuddin M, Petersen J, Mohammad A. Inactivation and modification of superoxide dismutase by glyoxal: prevention by antibodies. Biochimie 2007;89:311–8.PubMedCrossrefGoogle Scholar

  • 21.

    Kawaguchi M, Shibata N, Horiuchi S, Kobayashi M. Glyoxal inactivates glutamate transporter-1 in cultured rat astrocytes. Neuropathology 2005;25:27–36.PubMedGoogle Scholar

  • 22.

    Pedchenko VK, Chetyrkin SV, Chuang P, Ham AJ, Saleem MA, Mathieson PW, et al. Mechanism of perturbation of integrin-mediated cell-matrix interactions by reactive carbonyl compounds and its implication for pathogenesis of diabetic nephropathy. Diabetes 2005;54:2952–60.PubMedCrossrefGoogle Scholar

  • 23.

    Senolt L, Braun M, Vencovsky J, Sedova L, Pavelka K. Advanced glycation end-product pentosidine is not a relevant marker of disease activity in patients with rheumatoid arthritis. Physiol Res 2007;56:771–7.PubMedGoogle Scholar

  • 24.

    Busch M, Franke S, Wolf G, Brandstadt A, Ott U, Gerth J, et al. The advanced glycation end product Nε-carboxymethyllysine is not a predictor of cardiovascular events and renal outcomes in patients with type 2 diabetic kidney disease and hypertension. Am J Kidney Dis 2006;48:571–9.CrossrefGoogle Scholar

  • 25.

    Cotham WE, Metz TO, Ferguson PL, Brock JW, Hinton DJ, Thorpe SR, et al. Proteomic analysis of arginine adducts on glyoxal-modified ribonuclease. Mol Cell Proteomics 2004;3:1145–53.PubMedCrossrefGoogle Scholar

  • 26.

    Kawamura S, Chijiiwa Y, Minematsu T, Fukamizo T, Varum KM, Torikata T. The role of Arg114 at subsites E and F in reactions catalyzed by hen egg-white lysozyme. Biosci Biotechnol Biochem 2008;72:823–32.Google Scholar

  • 27.

    Verzijl N, DeGroot J, Thorpe SR, Bank RA, Shaw JN, Lyons TJ, et al. Effect of collagen turnover on the accumulation of advanced glycation end products. J Biol Chem 2000;275: 39027–31.Google Scholar

  • 28.

    Price RG, Spiro RG. Studies on the metabolism of the renal glomerular basement membrane. Turnover measurements in the rat with the use of radiolabeled amino acids. J Biol Chem 1977;252:8597–602.Google Scholar

  • 29.

    Thornalley PJ. Glyoxalase I – structure, function and a critical role in the enzymatic defence against glycation. Biochem Soc Trans 2003;31:1343–8.CrossrefPubMedGoogle Scholar

  • 30.

    Szwergold BS, Howell S, Beisswenger PJ. Human fructosamine-3-kinase: purification, sequencing, substrate specificity, and evidence of activity in vivo. Diabetes 2001;50: 2139–47.PubMedCrossrefGoogle Scholar

  • 31.

    Schleicher ED, Wagner E, Nerlich AG. Increased accumulation of the glycoxidation product Nε-(carboxymethyl)lysine in human tissues in diabetes and aging. J Clin Invest 1997;99:457–68.CrossrefGoogle Scholar

  • 32.

    Nakamura S, Tachikawa T, Tobita K, Aoyama I, Takayama F, Enomoto A, et al. An inhibitor of advanced glycation end product formation reduces Nε-(carboxymethyl)lysine accumulation in glomeruli of diabetic rats. Am J Kidney Dis 2003;41(3 Suppl 1):S68–71.CrossrefGoogle Scholar

  • 33.

    Zent R, Yan X, Su Y, Hudson BG, Borza DB, Moeckel GW, et al. Glomerular injury is exacerbated in diabetic integrin α1-null mice. Kidney Int 2006;70:460–70.Google Scholar

  • 34.

    Wendt T, Tanji N, Guo J, Hudson BI, Bierhaus A, Ramasamy R, et al. Glucose, glycation, and RAGE: implications for amplification of cellular dysfunction in diabetic nephropathy. J Am Soc Nephrol 2003;14:1383–95.CrossrefPubMedGoogle Scholar

  • 35.

    Glenn JV, Stitt AW. The role of advanced glycation end products in retinal ageing and disease. Biochim Biophys Acta 2009;1790:1109–16.Google Scholar

  • 36.

    Thornalley PJ, Battah S, Ahmed N, Karachalias N, Agalou S, Babaei-Jadidi R, et al. Quantitative screening of advanced glycation endproducts in cellular and extracellular proteins by tandem mass spectrometry. Biochem J 2003;375:581–92.Google Scholar

  • 37.

    Duran-Jimenez B, Dobler D, Moffatt S, Rabbani N, Streuli CH, Thornalley PJ, et al. Advanced glycation end products in extracellular matrix proteins contribute to the failure of sensory nerve regeneration in diabetes. Diabetes 2009;58:2893–903.CrossrefGoogle Scholar

  • 38.

    Pozzi A, Zent R, Chetyrkin S, Borza C, Bulus N, Chuang P, et al. Modification of collagen IV by glucose or methylglyoxal alters distinct mesangial cell functions. J Am Soc Nephrol 2009;20:2119–25.CrossrefPubMedGoogle Scholar

  • 39.

    Kislinger T, Fu C, Huber B, Qu W, Taguchi A, Du Yan S, et al. Nε-(carboxymethyl)lysine adducts of proteins are ligands for receptor for advanced glycation end products that activate cell signaling pathways and modulate gene expression. J Biol Chem 1999;274:31740–9.Google Scholar

  • 40.

    Heizmann CW. The mechanism by which dietary AGEs are a risk to human health is via their interaction with RAGE: arguing against the motion. Mol Nutr Food Res 2007;51:1116–9.PubMedCrossrefGoogle Scholar

  • 41.

    Ostendorp T, Leclerc E, Galichet A, Koch M, Demling N, Weigle B, et al. Structural and functional insights into RAGE activation by multimeric S100B. EMBO J 2007;26:3868–78.CrossrefGoogle Scholar

  • 42.

    Toure F, Zahm JM, Garnotel R, Lambert E, Bonnet N, Schmidt AM, et al. Receptor for advanced glycation end-products (RAGE) modulates neutrophil adhesion and migration on glycoxidated extracellular matrix. Biochem J 2008;416:255–61.Google Scholar

  • 43.

    Ban CR, Twigg SM. Fibrosis in diabetes complications: pathogenic mechanisms and circulating and urinary markers. Vasc Health Risk Manag 2008;4:575–96.Google Scholar

  • 44.

    Mott JD, Khalifah RG, Nagase H, Shield CF 3rd, Hudson JK, Hudson BG. Nonenzymatic glycation of type IV collagen and matrix metalloproteinase susceptibility. Kidney Int 1997;52:1302–12.PubMedCrossrefGoogle Scholar

  • 45.

    Sell DR, Biemel KM, Reihl O, Lederer MO, Strauch CM, Monnier VM. Glucosepane is a major protein cross-link of the senescent human extracellular matrix. Relationship with diabetes. J Biol Chem 2005;280:12310–5.Google Scholar

  • 46.

    Anderson SS, Wu K, Nagase H, Stettler-Stevenson WG, Kim Y, Tsilibary EC. Effect of matrix glycation on expression of type IV collagen, MMP-2, MMP-9 and TIMP-1 by human mesangial cells. Cell Adhes Commun 1996;4:89–101.PubMedGoogle Scholar

  • 47.

    Rosenthal AK, Gohr CM, Mitton E, Monnier V, Burner T. Advanced glycation end products increase transglutaminase activity in primary porcine tenocytes. J Investig Med 2009;57:460–6.PubMedGoogle Scholar

  • 48.

    Turk BE, Huang LL, Piro ET, Cantley LC. Determination of protease cleavage site motifs using mixture-based oriented peptide libraries. Nat Biotechnol 2001;19:661–7.PubMedCrossrefGoogle Scholar

  • 49.

    Khoshnoodi J, Sigmundsson K, Cartailler JP, Bondar O, Sundaramoorthy M, Hudson BG. Mechanism of chain selection in the assembly of collagen IV: a prominent role for the α2 chain. J Biol Chem 2006;281:6058–69.Google Scholar

  • 50.

    Howard EW, Benton R, Ahern-Moore J, Tomasek JJ. Cellular contraction of collagen lattices is inhibited by nonenzymatic glycation. Exp Cell Res 1996;228:132–7.Google Scholar

  • 51.

    Makino H, Shikata K, Hironaka K, Kushiro M, Yamasaki Y, Sugimoto H, et al. Ultrastructure of nonenzymatically glycated mesangial matrix in diabetic nephropathy. Kidney Int 1995;48:517–26.CrossrefPubMedGoogle Scholar

  • 52.

    Fathima NN, Madhan B, Rao JR, Nair BU, Ramasami T. Interaction of aldehydes with collagen: effect on thermal, enzymatic and conformational stability. Int J Biol Macromol 2004;34:241–7.CrossrefPubMedGoogle Scholar

  • 53.

    Metz TO, Alderson NL, Chachich ME, Thorpe SR, Baynes JW. Pyridoxamine traps intermediates in lipid peroxidation reactions in vivo: evidence on the role of lipids in chemical modification of protein and development of diabetic complications. J Biol Chem 2003;278:42012–9.Google Scholar

  • 54.

    Onorato JM, Jenkins AJ, Thorpe SR, Baynes JW. Pyridoxamine, an inhibitor of advanced glycation reactions, also inhibits advanced lipoxidation reactions. Mechanism of action of pyridoxamine. J Biol Chem 2000;275:21177–84.Google Scholar

  • 55.

    Lewis EJ, Greene T, Spitalewiz S, Blumenthal S, Berl T, Hunsicker LG, et al. Pyridorin in type 2 diabetic nephropathy. J Am Soc Nephrol 2012;23:131–6.Google Scholar

About the article

Corresponding author: Dr. Paul Voziyan, Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, S-3223 MCN, 1161 21st Avenue South, Nashville, TN 37232–2372, USA, Phone: +1-615-322-3352, Fax: +1-615-343-7156, E-mail:

Received: 2012-11-27

Accepted: 2013-02-07

Published Online: 2013-03-13

Published in Print: 2014-01-01

Citation Information: Clinical Chemistry and Laboratory Medicine, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2012-0818.

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