(A) X-ray structure of tricomplex of RMC-4998, K-Ras(G12C), and CypA (PDB: 8G9P). K-Ras residues involved in binding RMC-4998 are colored in blue. Negatively charged K-Ras residues involved in binding CypA are colored in orange.
(B) Sequence alignment of Ras-family GTPases K-Ras, M-Ras, R-Ras, and Rheb. Residues mediating drug resistance to RMC-4998 are highlighted in light orange (mild effect) and orange (strong effect).
(C) Immunoblot of HeLa cells transiently overexpressing K-Ras or K-Ras(G12C). HeLa cells were transiently transfected, treated with different concentration ofRMC-6291 for 3 h, and blotted for Ras.
(D) Immunoblot of HeLa cells transiently overexpressing M-Ras or M-Ras(G22C). HeLa cells were transiently transfected, treated with different concentration ofRMC-6291 for 3 h, and blotted for M-Ras.
(E) Immunoblot of HeLa cells transiently overexpressing R-Ras1 or R-Ras1(G38C). HeLa cells were transiently transfected, treated with different concentration ofRMC-6291 for 3 h, and blotted for R-Ras.
(F) Immunoblot of HeLa cells transiently overexpressing Rheb or Rheb(R15C). HeLa cells were transiently transfected, treated with different concentration of RMC-6291 for 3 h, and blotted for Rheb. Data are representative of three independent experiments.