title: A novel procedure for the assessment of the antioxidant capacity of food components.
abstract: Carbonylation, an oxidative modification of the amino group of arginine and lysine residues caused by reactive oxygen species, has emerged as a new type of oxidative damage. Protein carbonylation has been shown to exert adverse effects on various protein functions. Recently, the role of food components in the attenuation of oxidative stress has been the focus of many studies. Most of these studies focused on the chemical properties of food components. However, it is also important to determine their effects on protein functions via post-translational modifications. In this study, we developed a novel procedure for evaluating the antioxidant capacity of food components. Hydrogen peroxide (H2O2)-induced protein carbonylation in HL-60 cells was quantitatively analyzed by using fluorescent dyes (Cy5-hydrazide dye and IC3-OSu dye), followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorescence determination. Among a panel of food components tested, quinic acid, kaempferol, saponin, squalene, trigonelline, and mangiferin were shown to be capable of suppressing protein carbonylation in HL-60 cells. Our results demonstrated that this fluorescence labeling/SDS-PAGE procedure allows for the detection of oxidative stress-induced protein carbonylation with high sensitivity and quantitative accuracy. This method should be useful for the screening of new antioxidant food components as well as the analysis of their suppression mechanism.
authors.affiliation: Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, Miyazaki, Miyazaki, 889-2192, Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Yoshimura
authors.affiliation: Department of Biochemistry and Applied Biosciences, University of Miyazaki, Miyazaki, Miyazaki, 889-2192, Japan.
authors.forename: Mai
authors.initials: M
authors.lastname: Harashima
authors.affiliation: Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, Miyazaki, Miyazaki, 889-2192, Japan; Department of Biochemistry and Applied Biosciences, University of Miyazaki, Miyazaki, Miyazaki, 889-2192, Japan.
authors.forename: Katsuhisa
authors.initials: K
authors.lastname: Kurogi
authors.affiliation: Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, Miyazaki, Miyazaki, 889-2192, Japan; Department of Biochemistry and Applied Biosciences, University of Miyazaki, Miyazaki, Miyazaki, 889-2192, Japan.
authors.forename: Masahito
authors.initials: M
authors.lastname: Suiko
authors.affiliation: Department of Pharmacology, College of Pharmacy and Pharmaceutical Sciences, University of Toledo, Toledo, OH, 43614, USA.
authors.forename: Ming-Cheh
authors.initials: MC
authors.lastname: Liu
authors.affiliation: Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, Miyazaki, Miyazaki, 889-2192, Japan; Department of Biochemistry and Applied Biosciences, University of Miyazaki, Miyazaki, Miyazaki, 889-2192, Japan. Electronic address: ysakaki@cc.miyazaki-u.ac.jp.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Sakakibara
mesh_terms: D000470:Alkaloids / Q000737:chemistry / Q000494:pharmacology; D000975:Antioxidants / Q000737:chemistry* / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000737:chemistry; D005502:Food; D018922:HL-60 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors / Q000494:pharmacology; D044949:Kaempferols / Q000737:chemistry / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects; D011506:Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D011801:Quinic Acid / Q000737:chemistry / Q000494:pharmacology; D012503:Saponins / Q000737:chemistry / Q000494:pharmacology; D013185:Squalene / Q000737:chemistry / Q000494:pharmacology; D044004:Xanthones / Q000737:chemistry / Q000494:pharmacology
chemical_list: D000470:Alkaloids; D000975:Antioxidants; D005456:Fluorescent Dyes; D044949:Kaempferols; D011506:Proteins; D012503:Saponins; D044004:Xanthones; D011801:Quinic Acid; C013592:mangiferin; C009560:trigonelline; C006552:kaempferol; D013185:Squalene; D006861:Hydrogen Peroxide
keywords: Antioxidant; Carbonylation; Fluorescence labeling electrophoresis; Oxidative stress