Fig. 2.
Combined assays of the autokinase, phosphoryltransfer, and phosphatase activities mediated by truncated WalK+ or WalKT222A. Reactions were performed at 25°C in reaction mixtures containing Mg2+ or Ca2+ buffer as described in Experimental procedures. Autophosphorylation reactions containing 2.0 μM of purified (A) (N)-Sumo-ΔN35-WalK+ or (B) (N)-Sumo-ΔN35-WalKT222A protein proceeded for 30 min before (N)-His-WalR (final concn = 6.6 μM) was added without removal of ATP (t=0). The experiment was repeated >3 times, and representative phosphorimages of time courses are shown.