Conformational change of E.coli sulfurtransferase YgaP upon SCN- in intact native membrane revealed by fluorescence lifetime and anisotropy  

作　　者：Simeng Wang Yanan Zhang Longhua Zhang Min Zhang Changlin Tian 

机构地区：[1]School of Life Science, Anhui University, Hefei 230039, China [2]National Laboratory of Physical Sciences at Microscale and School of Life Science, University of Science and Technology of China, Hefei 230027, China [3]High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei 230031, China

出　　处：《Chinese Chemical Letters》2018年第10期1513-1516,共4页中国化学快报（英文版）

基　　金：supported by the National Key R&D Program of China (Nos. 2016YFA0400900, 2017YFA0505300);the Instrument Developing Project of the Chinese Academy of Sciences (No. YZ201564)

摘　　要：Fluorescence lifetime and anisotropy has become a prevalent tool to detect the structure change and motility property of proteins. YgaP is the only membrane-integrated rhodanese in E. coli. The sulfur transfer process has been characterized by various studies. However, the mechanism of the outward transportation of SCN-remains unclear. In this work, we examined the fluorescence lifetime and anisotropy of site-specific incorporated unnatural amino acid 7-HC to study the conformational change of YgaP upon SCN-binding. We also compared the fluorescence changes between detergent-wrapped environment in DPC and intact native membrane environment in SMA. Our results suggested the presence of at least two different conformations in YgaP protein. Both the residues in the middle of TMH2 and the residues near extracellular side play important roles in the binding and/or output of SCN-. SMA is a good material to reflect the in situ conformation changes of protein than micelles.Fluorescence lifetime and anisotropy has become a prevalent tool to detect the structure change and motility property of proteins. YgaP is the only membrane-integrated rhodanese in E. coli. The sulfur transfer process has been characterized by various studies. However, the mechanism of the outward transportation of SCN^- remains unclear. In this work, we examined the fluorescence lifetime and anisotropy of site-specific incorporated unnatural amino acid 7-HC to study the conformational change of YgaP upon SCN^- binding. We also compared the fluorescence changes between detergent-wrapped environment in DPC and intact native membrane environment in SMA. Our results suggested the presence of at least two different conformations in YgaP protein. Both the residues in the middle of TMH2 and the residues near extracellular side play important roles in the binding and/or output of SCN^-. SMA is a good material to reflect the in situ conformation changes of protein than micelles.

关 键 词：Rhodanese YgaP Fluorescence lifetime and anisotropy SMA Conformation change 

分 类 号：O657.3[理学—分析化学] Q51[理学—化学]