Products/Services Used | Details | Operation |
---|---|---|
Bacterial Expression> | A synthetic gene encoding KRAS4B residues 1–173 (with C118S mutation) optimized for Escherichia coli expression (GenScript) was cloned into a pET-28 vector and expressed in E. coli (BL21) (residue 1–173, with C118S mutation) | Get A Quote |
Cancer cells bearing distinct KRAS mutations exhibit variable sensitivity to SHP2 inhibitors (SHP2i). Here we show that cells harboring KRAS Q61H are uniquely resistant to SHP2i, and investigate the underlying mechanisms using biophysics, molecular dynamics, and cell-based approaches. Q61H mutation impairs intrinsic and GAP-mediated GTP hydrolysis, and impedes activation by SOS1, but does not alter tyrosyl phosphorylation. Wild-type and Q61H-mutant KRAS are both phosphorylated by Src on Tyr32 and Tyr64 and dephosphorylated by SHP2, however, SHP2i does not reduce ERK phosphorylation in KRAS Q61H cells. Phosphorylation of wild-type and Gly12-mutant KRAS, which are associated with sensitivity to SHP2i, confers res... More