文献类型：期刊文献

英文题名：Proteome-Wide Deconvolution of Drug Targets and Binding Sites by Lysine Reactivity Profiling

作者：Ruan, Chengfei[1,2];Zhou, Jiahua[1,2];Li, Zhouxian[1,3];Li, Kejia[1,2];Fang, Zheng[1,2];Zhang, Xiaolei[1];Ye, Mingliang[1]

机构：[1]Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China;[2]Univ Chinese Acad Sci, Beijing 100049, Peoples R China;[3]East China Univ Sci & Technol, Dept Chem & Mol Engn, Shanghai Key Lab Funct Mat Chem, Shanghai 200237, Peoples R China

年份：2022

卷号：94

期号：7

起止页码：3352

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20220911725515);WOS:【SCI-EXPANDED(收录号:WOS:000758083900001)】；

基金：This work was supported, in part, by funds from the National Key Research and Development Program of China (2020YFE0202200, 2017YFA0505003), the National Natural Science Foundation of China (92153302, 22137002), the LiaoNing Revitalization Talents Program, the Innovation Program of Science and Research from the DICP, CAS (DICP I201935, DICP & QIBEBT UN201802), and the Innovation Academy for Precision Measurement Science and Technology, CAS.

语种：英文

外文关键词：Probes - Amino acids - Proteins

摘要：Recently, numerous efforts have been devoted to identifying drug targets and binding sites in complex proteomes, which is of great importance in modern drug discovery. In this study, we developed a robust lysine reactivity profiling method to systematically study drug-binding targets and binding sites at the proteome level. This method is based on the principle that binding of a drug to a specific region of target proteins will change the reactivity of lysine residues that are located at this region, and these changes can be detected with an enrichable and lysine reactive probe. Coupled with data-independent acquisition (DIA), the known target proteins and corresponding binding sites were successfully revealed from K562 cell lysates for three model drugs: geldanamycin, staurosporine, and dasatinib. In addition, the druginduced conformational changes of certain targets were also revealed by our method during the screening of staurosporine. The screening sensitivity of our method revealed from the screening of stuarosporine and dasatinib was comparable with that of thermal proteome profiling (TPP) or machine learning-based limited proteolysis (LiP-Quant). Overall, 21 and 4 kinase targets, including adenosine 5'-triphosphate (ATP)-binding targets, were identified for staurosporine and dasatinib in K562 cell lysates, respectively. We found that target proteins identified by TPP, LiP-Quant, and our method were complementary, emphasizing that the development of new methods that probe different properties of proteins is of great importance in drug target deconvolution. We also envision further applications of our method in proteome-wide probing multiple events that involve lysine reactivity changes.