Fluorescence lifetime based distance measurement illustrates conformation changes of PYL10-CL2 upon ABA binding in solution state

作     者：Peng Zhou Pei Lv Lu Yu Sanling Liu Longhua Zhang Changlin Tian 

作者机构：High Magnetic Field Laboratory Chinese Academy of Sciences and School of Life Sciences University of Science and Technology of China 

出 版 物：《Chinese Chemical Letters》 (中国化学快报（英文版）)

年 卷 期：2019年第30卷第5期

页      面：1067-1070页

核心收录：

学科分类：07[理学] 0703[理学-化学] 

基　　金：supported by National Key R&D Program of China(Nos.2017YFA0505300,2016YFA0400900) the Instrument Developing Project of the Chinese Academy of Sciences(No.YZ201564) the National Natural Science Foundation of China(Nos.U1832181,31670776,31500611) 

主　　题：Fluorescence lifetime Anisotropy Unnatural amino acid PYL10 Distance measurement 

摘      要：Forster resonance energy transfer(FRET)is a widely used distance measurement method to illustrate protein conformational *** FRET method relies on the distance between donor and acceptor,as well as the labelling efficiency,the size and the properties of the ***,we labelled a pair of small fluorophores and calculated the energy transferred efficiency through fluorescence lifetime analysis,which can provide more reliable distance measurement than intensity *** donor fluorophore,7-hydroxycoumarin-4-yl-ethylglycine(HC),was genetically incorporated into specific sites of PYL10,obtaining complete labelling *** acceptor fluorophore,Alexa488,was labelled through the disulfide bond,whose labelling efficiency was estimated through both absorption peaks and lifetime *** lifetime and anisotropy analysis showed ABA-induced local conformation changes and dynamics of several HC incorporation sites of *** lifetime-based FRET distance measurement illustrated the conformation changes of PYL10 with or without ABA application,which is consistent with the previously reported crystal structures.