Recombinant Human CDC42 fused with a histidine tag in the amino-terminus was expressed in E. coli.
The constitutively active form of human Cdc42 protein contains a glutamine to leucine substitution at residue 61. The common name for this mutant is Cdc42(Q61L) or L61Cdc42. The leucine substitution prevents endogenous and GAP-stimulated GTPase activity of Cdc42, hence the protein is always in the active, GTP-bound, state. A similar mutation in Ras (Ras(Q61L)) behaves as an oncogenic mutation.
2 mM Tris pH 7.6, 0.5 mM MgCl2, 0.5% sucrose and 0.1% dextran.
Cdc42 effector and GAP identification and binding studies; Activation of Cdc42 effectors in vitro and in vivo; Stimulation of Cdc42 phenotype by microinjection into cells