蛋白质组
蛋白质组学
巴基斯坦卢比
化学
生物
分子生物学
生物化学
丙酮酸激酶
基因
新陈代谢
糖酵解
作者
Ikuko Nagasawa,Makoto Muroi,Masahito Kawatani,Tomokazu Ohishi,Seigo Ohba,Manabu Kawada,Hiroyuki Osada
标识
DOI:10.1016/j.chembiol.2019.11.010
摘要
Summary
The cellular thermal shift assay (CETSA) has recently been devised as a label-free method for target validation of small compounds and monitoring the thermal stabilization or destabilization of proteins due to binding with the compound. Herein, we developed a modified method by combining the CETSA and proteomics analysis based on 2D gel electrophoresis, namely 2DE-CETSA, to identify the thermal stability-shifted proteins by binding with a new compound. We applied the 2DE-CETSA for analysis of a target-unknown compound, NPD10084, which exerts anti-proliferative activity against colorectal cancer cells in vitro and in vivo, and identified pyruvate kinase muscle isoform 2 (PKM2) as a candidate target protein. Interestingly, NPD10084 interrupted protein-protein interactions between PKM2 and β-catenin or STAT3, with subsequent suppression of downstream signaling. We thus demonstrate that our 2DE-CETSA method is applicable for identification of target compounds discovered by phenotypic screening.
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