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Analysis of hydroxyl radical‐induced oxidation process of glucagon by reversed‐phase HPLC and ESI‐MS/MS
Author(s) -
Ichiba Hideaki,
Ogawa Tomoko,
Yajima Takehiko,
Fukushima Takeshi
Publication year - 2009
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1222
Subject(s) - chemistry , chromatography , high performance liquid chromatography , phase (matter) , process (computing) , organic chemistry , computer science , operating system
Abstract Structural modification of a polypeptide hormone, glucagon, by a hydroxyl radical in vitro was investigated by reversed‐phase high‐performance liquid chromatography (RP‐HPLC), and the oxidized site of glucagon was detected by electrospray ionization tandem mass spectrometry (ESI‐MS/MS). It was shown that 27 methionine (Met) was oxidized to 27 Met sulfoxide by hydroxyl radical, and the production rate of 27 Met sulfoxide was faster than that by hydrogen peroxide. In addition, production of 27 Met sulfoxide enantiomer was confirmed by RP‐HPLC analysis. cAMP production in a HepG2 cell induced by 27 Met sulfoxide glucagon was reduced to approximately 75% as compared with that induced by the native form of glucagon. Copyright © 2009 John Wiley & Sons, Ltd.