The role of alpha- and epsilon-amino groups in the glycation-mediated cross-linking of gammaB-crystallin. Study of three site-directed mutants.

The Journal of Biological Chemistry
H R ZhaoE C Abraham

Abstract

In the previous report we demonstrated that gammaB-crystallin is glycated predominantly at the N-terminal alpha-amino group (Casey, E. B., Zhao, H. R., and Abraham, E. C. (1995) J. Biol. Chem. 270, 20781-20786). To investigate the possible role of alpha- and epsilon-amino groups of gammaB-crystallin in glycation-mediated cross-linking, Lys-2 or Lys-163, or both, were mutated to threonine by site-directed mutagenesis in bovine gammaB-crystallin cDNA. Wild type and mutant gammaB-crystallins were expressed in Escherichia coli cells. Cross-linking studies were performed by incubating wild type and mutant gammaB-crystallins with glyceraldehyde, ribose, and galactose followed by SDS-polyacrylamide gel electrophoresis under reducing conditions. When both of the lysines of gammaB-crystallin were mutated to threonines (gammaB-K2T/K163T), the quantity of cross-linked products was greatly reduced, indicating that, despite the fact that the alpha-amino group is a major glycated site, epsilon-amino groups play a predominant role in cross-linking. Therefore, cross-linking ability depends not only upon the level of glycation but also upon which amino group is glycated. Steric hindrance may decrease the cross-linking ability of the alpha-amino...Continue Reading

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Citations

Aug 16, 2000·Proceedings of the National Academy of Sciences of the United States of America·Q YangY S Kanwar
Jun 17, 1998·Experimental Eye Research·E SpoerlT Seiler
Dec 26, 2016·International Journal of Biological Macromolecules·Susmitnarayan ChaudhurySwagata Dasgupta
Dec 29, 2016·International Journal of Biological Macromolecules·Abdul Rouf MirSafia Habib
Mar 13, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yang YiHong-Xun Wang
Sep 13, 2002·The Journal of Biological Chemistry·Hiroshi SakamotoTakashi Tsuruo

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