A Chemical-Genetic Approach to Generate Selective Covalent Inhibitors of Protein Kinases

作者：Kung Alvin; Schimpl Marianne; Ekanayake Arunika; Chen Ying Chu; Overman Ross; Zhang Chao^*

来源：ACS Chemical Biology, 2017, 12(6): 1499-1503.

DOI：10.1021/acschembio.6b01083

Although a previously developed bump-hole approach has proven powerful in generating specific inhibitors for mapping functions of protein kinases, its application is limited by the intolerance of the large-to-small mutation by certain kinases and the inability to control two kinases separately in the same cells. Herein, we describe the development of an alternative chemical-genetic approach to overcome these limitations. Our approach features the use of an engineered cysteine residue at a particular position as a reactive feature to sensitize a kinase of interest to selective covalent blockade by electrophilic inhibitors and is thus termed the Ele-Cys approach. We successfully applied the Ele-Cys approach to identify selective covalent inhibitors of a receptor tyrosine kinase EphB1 and solved cocrystal structures to determine the mode of covalent binding. Importantly, the Ele-Cys and bump-hole approaches afforded orthogonal inhibition of two distinct kinases in the cell, opening the door to their combined use in the study of multikinase signaling pathways.

• 出版日期2017-6