Molecular dynamics simulations reveal the activation mechanism of mutations G12V and Q61L of Cdc42

Abstract Cell division control protein 42 homolog (Cdc42), which contributes to multiple cellular processes including cell proliferation and migration, is a potential target for cancer therapy, especially in the intervention of tumor migration. Cdc42's mutants G12V and Q61L are discovered constitutively active, and the overexpression of them exhibits oncogenic activities. Here, using molecular dynamics (MD) simulations and dynamic analysis, we illustrated the activation mechanism of Cdc42 G12V and Cdc42 Q61L . Without GAP, the two mutations differently elicited state transition from the wild‐type's open “inactive” state 1 to the closed “active” state 2, induced by the introduction of a newly formed water‐mediated T35‐γ‐phosphate hydrogen bond in G12V system and the additional hydrophobic interactions between L61 and T35 together with the direct T35‐γ‐phosphate hydrogen bond in Q61L system. When binding with GAP, both mutations weakened the hydrogen bond interactions between Cdc42‐GTP and GAP's finger loop, and disturbed the catalytically competent organizations of GAP's catalytic R305/R306 and Cdc42's Q61, thereby impairing the GAP‐mediated GTP hydrolysis. Our findings first reveal the activation mechanism of Cdc42's G12V and Q61L mutants on a molecular basis, which provide new insights into the structural and dynamical characteristics of Cdc42 and its mutants and can be exploited in the further development of novel therapies targeting Cdc42‐related cancers.