Identification of glucose-derived cross-linking sites in ribonuclease A.

Journal of Proteome Research
Zhenyu DaiVincent M Monnier

Abstract

The accumulation of glycation derived cross-links has been widely implicated in extracellular matrix damage in aging and diabetes, yet little information is available on the cross-linking sites in proteins and the intra- versus intermolecular character of cross-linking. Recently, glucosepane, a 7-membered heterocycle formed between lysine and arginine residues, has been found to be the single major cross-link known so far to accumulate during aging. As an approach toward identification of glucose derived cross-linking sites, we have preglycated ribonuclease A first for for 14 days with 500 mM glucose, followed by a 4-week incubation in absence of glucose. MALDI-TOF analysis of tryptic digests revealed the presence of Amadori products (Delta m/ z = 162) at K1, K7, K37 and K41, in accordance with previous studies. In addition, K66, K98 and K104 were also modified by Amadori products. Intramolecular glucosepane cross-links were observed at K41-R39 and K98-R85. Surprisingly, the only intermolecular cross-link observed was the 3-deoxyglucosone-derived DODIC at K1-R39. The identity of cross-linked peptides was confirmed by sequencing with tandem mass spectrometry. Recombinant ribonuclease A mutants R39A, R85A, and K91A were produced,...Continue Reading

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Citations

Sep 16, 2008·Analytical and Bioanalytical Chemistry·Yuyuan LiJoel A Dain
Jan 10, 2012·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Brittany LawEdie C Goldsmith
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Feb 4, 2012·Journal of Proteomics·María Ramírez-BooJean-Charles Sanchez
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Jan 10, 2012·Gerontology·David R Sell, Vincent M Monnier
Jun 22, 2013·Clinical Chemistry and Laboratory Medicine : CCLM·Vincent M MonnierSaul Genuth
Mar 27, 2021·Annual Review of Food Science and Technology·Yan ZhangShuo Wang
Jul 13, 2021·Mass Spectrometry Reviews·Giovanni RenzoneAndrea Scaloni
Oct 9, 2018·Journal of Agricultural and Food Chemistry·Alexander KlausMarcus A Glomb

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