The Maillard or browning reaction between sugar and protein contributes to the increased chemical modification and cross-linking of long-lived tissue proteins in diabetes. To evaluate the role of glycation and oxidation in these reactions, we have studied the effects of oxidative and antioxidative conditions and various types of inhibitors on the reaction of glucose with rat tail tendon collagen in phosphate buffer at physiological pH and temperature. The chemical modifications of collagen that were measured included fructoselysine, the glycoxidation products Nε-(carboxymethyl)lysine and pentosidine and fluorescence. Collagen cross-linking was evaluated by analysis of cyanogen bromide peptides using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by changes in collagen solubilization on treatment with pepsin or sodium dodecylsulfate. Although glycation was unaffected, formation of glycoxidation products and cross-linking of collagen were inhibited by antioxidative conditions. The kinetics of formation of glycoxidation products proceeded with a short lag phase and were independent of the amount of Amadori adduct on the protein, suggesting that autoxidative degradation of glucose was a major contributor to glycoxidation and cross-linking reactions. Chelators, sulfhydryl compounds, antioxidants, and aminoguanidine also inhibited formation of glycoxidation products, generation of fluorescence, and cross-linking of collagen without significant effect on the extent of glycation of the protein. We conclude that autoxidation of glucose or Amadori compounds on protein plays a major role in the formation of glycoxidation products and cross-liking of collagen by glucose in vitro and that chelators, sulfhydryl compounds, antioxidants, and aminoguanidine act as uncouplers of glycation from subsequent glycoxidation and cross-linking reactions.
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Original Articles|
May 01 1994
Glycation, Glycoxidation, and Cross-Linking of Collagen by Glucose: Kinetics, Mechanisms, and Inhibition of Late Stages of the Maillard Reaction
Min-Xin Fu;
Min-Xin Fu
Department of Chemistry and Biochemistry, University of South Carolina
Columbia
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Kevin J Wells-Knecht;
Kevin J Wells-Knecht
Department of Chemistry and Biochemistry, University of South Carolina
Columbia
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James A Blackledge;
James A Blackledge
Department of Chemistry and Biochemistry, University of South Carolina
Columbia
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Thorpe J Lyons;
Thorpe J Lyons
Division of Endocrinology, Diabetes, and Metabolism, Medical University of South Carolina
Charleston, South Carolina
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Suzanne R Thorpe;
Suzanne R Thorpe
Department of Chemistry and Biochemistry, University of South Carolina
Columbia
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John W Baynes
John W Baynes
Department of Chemistry and Biochemistry, University of South Carolina
Columbia
School of Medicine, University of South Carolina
Columbia
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Address correspondence and reprint requests to Dr. John W. Baynes, Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC 29208
Diabetes 1994;43(5):676–683
Article history
Received:
July 23 1993
Revision Received:
January 13 1994
Accepted:
January 13 1994
PubMed:
8168645
Citation
Min-Xin Fu, Kevin J Wells-Knecht, James A Blackledge, Thorpe J Lyons, Suzanne R Thorpe, John W Baynes; Glycation, Glycoxidation, and Cross-Linking of Collagen by Glucose: Kinetics, Mechanisms, and Inhibition of Late Stages of the Maillard Reaction. Diabetes 1 May 1994; 43 (5): 676–683. https://doi.org/10.2337/diab.43.5.676
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