Work

Protocol to identify covalent inhibitors targeting RhoA Cys16

Year: 2026

Type: article

Abstract: Ras homolog family member A (RhoA) is a therapeutic hotspot in many cancers but remains undruggable. Here, we present a protocol for identifying covalent inhibitors targeting RhoA Cys16. We describe steps for the identification of lead compounds by activity-based protein profiling (ABPP) screening and the validation of engagement by mass spectrometry and chemical biology experiments. We then detail procedures for investigating the effects of the compounds on RhoA signaling in cancer cells and mouse colorectal cancer (CRC) models. For complete details on the use and execution of this protocol, please refer to Koo et al. 1

Source: STAR Protocols

DOI: 10.1016/j.xpro.2026.104494

Authors: Tin-Yan Koo, Chloe See-Hang Ho, Hillary Yui-Yan Yip, Clive Yik-Sham Chung

Institution: University of Hong Kong

Language: English

Topic: Protein Kinase Regulation and GTPase Signaling

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550