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Hydrogen peroxide-induced chlorophyll a bleaching in the cytochrome b 6 f complex: a simple and effective assay for stability of the complex in detergent solutions
Authors:Xiao-Bo Chen  Xiao-Hui Zhao  Yi Zhu  Yan-Dao Gong  Liang-Bi Li  Jian-Ping Zhang  Ting-Yun Kuang
Institution:(1) Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Nan Xincun 20, Xiangshan, 100093 Beijing, P.R. China;(2) State Key Laboratory for Structural Chemistry of Unstable and Stable Species, Institute of Chemistry, Chinese Academy of Sciences, 1st North Street, Zhongguancun, 100080 Beijing, P.R. China;(3) Department of Biological Sciences and Biotechnology, Tsinghua University, 100084 Beijing, P.R. China
Abstract:The instability of cytochrome b 6 f complex in detergent solutions is a well-known problem that has been studied extensively, but without finding a satisfactory solution. One of the important reasons can be short of the useful method to verify whether the complex suspended in different detergent is in an intact state or not. In this article, a simple and effective assay for stability of the complex was proposed based on the investigation on the different effects of the two detergents, n-octyl-β-d-glucopyranoside (OG) and dodecyl-β-d-maltoside (DDM), on the properties of the complex. DDM stabilizes the complex preparation more effectively whereas OG denatures the interactions of the heme groups and pigment molecules with the protein environment, leading to the bleaching of chlorophyll a induced by addition of hydrogen peroxide. The assay of the use of hydrogen peroxide to characterize the complex by studying the bleaching of chlorophyll induced by hydrogen peroxide and the peroxidase activity of the complex was discussed. This simple method will probably be useful to study the stability of the complex. Xiao-Bo Chen and Xiao-Hui Zhao contributed equally to this work.
Keywords:Cytochrome b                      6                      f            Detergent  Chlorophyll  Peroxidase activity  Hydrogen peroxide
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