title: Analysis of hydroxyl radical-induced oxidation process of glucagon by reversed-phase HPLC and ESI-MS/MS.
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.
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Toho University, 2-2-1 Miyama, Funabashi-shi, Chiba 274-8510, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Ichiba
authors.forename: Tomoko
authors.initials: T
authors.lastname: Ogawa
authors.forename: Takehiko
authors.initials: T
authors.lastname: Yajima
authors.forename: Takeshi
authors.initials: T
authors.lastname: Fukushima
mesh_terms: D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000242:Cyclic AMP / Q000378:metabolism; D005934:Glucagon / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D056945:Hep G2 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007501:Iron / Q000378:metabolism; D016014:Linear Models; D008715:Methionine / Q000378:metabolism; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species / Q000378:metabolism; D012451:Safrole / Q000031:analogs & derivatives / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013997:Time Factors
chemical_list: D017382:Reactive Oxygen Species; C036094:glucagon, Met-sulfoxide(27)-; D005934:Glucagon; D008715:Methionine; D006861:Hydrogen Peroxide; D000242:Cyclic AMP; D007501:Iron; D012451:Safrole; C005746:sulfoxide