Wang, Andrew, Pham, Helen, Lipps, Jennifer, Brittain, Scott, Harrington, Edmund, Wang, Yuan, King, Frederick, Russ, Carsten, Pan, Xuewen, Hoepfner, Dominic, Tallarico, John, Feng, Yan, Jain, Rishi, Schirle, Markus and Thomas, Jason (2019) Previously Uncharacterized Vacuolar-type ATPase Binding Site Discovered from Structurally Similar Compounds with Distinct Mechanisms of Action. ACS Chemical Biology, 14 (1). pp. 20-26. ISSN 15548937

Abstract

Using a comprehensive chemical genetics approach, we identified a member of the lignan natural product family, HTP-013, which exhibited significant cytotoxicity across various cancer cell lines. Correlation of compound activity across a panel of reporter gene assays suggested the vacuolar-type ATPase (v-ATPase) as a potential target for this compound. Additional cellular studies and a yeast haploinsufficiency screen strongly supported this finding. Competitive photoaffinity labeling experiments demonstrated that the ATP6V0A2 subunit of the v-ATPase complex binds directly to HTP-013, and further mutagenesis library screening identified resistance-conferring mutations in ATP6V0A2. The positions of these mutations suggest the molecule binds a novel pocket within the domain of the v-ATPase complex responsible for proton translocation. While other mechanisms of v-ATPase regulation have been described, such as dissociation of the complex or inhibition by natural products including bafilomycin A1 and concanamycin, this work provides detailed insight into a distinct binding pocket within the v-ATPase complex.

Item Type:	Article
Date Deposited:	19 Jun 2021 00:45
Last Modified:	19 Jun 2021 00:45
URI:	https://oak.novartis.com/id/eprint/34011