Identification of the site of glycation of human insulin

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

Research output: Contribution to journalArticle

46 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.
LanguageEnglish
Pages1323-1330
JournalPeptides
Volume17
Issue number8
Publication statusPublished - 1996

Fingerprint

Mass spectrometry
Insulin
Thermolysin
Sorbitol
Molecular mass
Disulfides
Trypsin
Desorption
Plasmas
Glucose
Degradation

Cite this

O'Harte, F., Hojrup, P., Barnett, CR., & Flatt, P. (1996). Identification of the site of glycation of human insulin. 17(8), 1323-1330.
O'Harte, Finbarr ; Hojrup, P ; Barnett, CR ; Flatt, Peter. / Identification of the site of glycation of human insulin. 1996 ; Vol. 17, No. 8. pp. 1323-1330.
@article{9d44c3074198403d870ba1bddcd251c9,
title = "Identification of the site of glycation of human insulin",
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.",
author = "Finbarr O'Harte and P Hojrup and CR Barnett and Peter Flatt",
year = "1996",
language = "English",
volume = "17",
pages = "1323--1330",
number = "8",

}

O'Harte, F, Hojrup, P, Barnett, CR & Flatt, P 1996, 'Identification of the site of glycation of human insulin', vol. 17, no. 8, pp. 1323-1330.

Identification of the site of glycation of human insulin. / O'Harte, Finbarr; Hojrup, P; Barnett, CR; Flatt, Peter.

Vol. 17, No. 8, 1996, p. 1323-1330.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of the site of glycation of human insulin

AU - O'Harte, Finbarr

AU - Hojrup, P

AU - Barnett, CR

AU - Flatt, Peter

PY - 1996

Y1 - 1996

N2 - 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.

AB - 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.

M3 - Article

VL - 17

SP - 1323

EP - 1330

IS - 8

ER -