Identification of the site of glycation of human insulin

Finbarr O'Harte, P Hojrup, CR Barnett, Peter Flatt

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51 Citations (Scopus)

Abstract

This study evaluates the nature of glycated human insulin formed following exposure to hyperglycemic conditions in vitro. Glycated insulin was purified by RP-HPLC and its molecular mass (5971.3 Da) determined by plasma desorption mass spectrometry (MS). The difference in mass (163.7 Da) from nonglycated insulin (5807.6 Da) corresponds to a single reduced glucose (glucitol) residue. Following reduction of insulin disulfide bridges, MS confirmed that the B-chain was glycated. Enzymatic digestions with trypsin, endoproteinase Glu-C, and thermolysin, followed by RP-HPLC and identification of fragments by MS, localized glycation to the B-chain (1-5) region. Electrospray tandem MS identified the site of glycation as the B-chain NH2-terminal Phe(1) residue. This was confirmed by automated Edman degradation with glycated human insulin. Copyright (C) 1996 Elsevier Science Inc.
Original languageEnglish
Pages (from-to)1323-1330
JournalPeptides
Volume17
Issue number8
Publication statusPublished - 1996

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