Global profiling of AMG510 modified proteins identified tumor suppressor KEAP1 as an off-target
Yini Wang,
Bowen Zhong,
Caixia Xu,
Dongdong Zhan,
Songhao Zhao,
Hongxing Wu,
Mingwei Liu,
Xiaoling Lan,
Danni Cai,
Qian Ding,
Biao Zheng,
Jiong Lan,
Qiang Lv,
Yi Wang,
Jun Qin
Affiliations
Yini Wang
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (The PHOENIX Center, Beijing), Beijing Institute of Lifeomics, Beijing 102206, China
Bowen Zhong
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (The PHOENIX Center, Beijing), Beijing Institute of Lifeomics, Beijing 102206, China
Caixia Xu
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (The PHOENIX Center, Beijing), Beijing Institute of Lifeomics, Beijing 102206, China
Dongdong Zhan
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (The PHOENIX Center, Beijing), Beijing Institute of Lifeomics, Beijing 102206, China
Songhao Zhao
Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
Hongxing Wu
Beijing Pineal Diagnostics Co., Ltd., Beijing 100195, China
Mingwei Liu
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (The PHOENIX Center, Beijing), Beijing Institute of Lifeomics, Beijing 102206, China
Xiaoling Lan
GenFleet Therapeutics Inc., Shanghai 201203, China
Danni Cai
GenFleet Therapeutics Inc., Shanghai 201203, China
Qian Ding
GenFleet Therapeutics Inc., Shanghai 201203, China
Biao Zheng
GenFleet Therapeutics Inc., Shanghai 201203, China
Jiong Lan
GenFleet Therapeutics Inc., Shanghai 201203, China
Qiang Lv
GenFleet Therapeutics Inc., Shanghai 201203, China
Yi Wang
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (The PHOENIX Center, Beijing), Beijing Institute of Lifeomics, Beijing 102206, China
Jun Qin
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (The PHOENIX Center, Beijing), Beijing Institute of Lifeomics, Beijing 102206, China; Corresponding author
Summary: KRAS inhibitor AMG510 covalently modifies the G12C residue and inactivates the KRAS/G12C function. Because there are many reactive cysteines in the proteome, it is important to characterize AMG510 on-target modification and off-targets. Here, we presented a streamlined workflow to measure abundant AMG510 modified peptides including that of KRAS/G12C by direct profiling, and a pan-AMG510 antibody peptide IP workflow to profile less abundant AMG510 off-targets. We identified over 300 off-target sites with three distinct kinetic patterns, expanding the AMG510 modified proteome involved in the nucleocytoplasmic transport, response to oxidative stress, adaptive immune system, and glycolysis. We found that AMG510 covalently modified cys339 of ALDOA and inhibited its enzyme activity. Moreover, AMG510 modified KEAP1 cys288 and induced NRF2 accumulation in the nuclear of NSCLC cells independent of KRAS/G12C mutation. Our study provides a comprehensive resource of protein off-targets of AMG510 and elucidates potential toxicological sideeffects for this covalent KRASG12C inhibitor.
