K stimulation of ATPase activity associated with the chloroplast inner envelope.

Studies were conducted to characterize ATPase activity associated with purified chloroplast inner envelope preparations from spinach (Spinacea oleracea L.) plants. Comparison of free Mg(2+) and Mg.ATP complex effects on ATPase activity revealed that any Mg(2+) stimulation of activity was likely a function of the use of the Mg.ATP complex as a substrate by the enzyme; free Mg(2+) may be inhibitory. In contrast, a marked (one- to twofold) stimulation of ATPase activity was noted in the presence of K(+). This stimulation had a pH optimum of approximately pH 8.0, the same pH optimum found for enzyme activity in the absence of K(+). K(+) stimulation of enzyme activity did not follow simple Michaelis-Menton kinetics. Rather, K(+) effects were consistent with a negative cooperativity-type binding of the cation to the enzyme, with the K(m) increasing at increasing substrate. Of the total ATPase activity associated with the chloroplast inner envelope, the K(+)-stimulated component was most sensitive to the inhibitors oligomycin and vanadate. It was concluded that K(+) effects on this chloroplast envelope ATPase were similar to this cation's effects on other transport ATPases (such as the plasmalemma H(+)-ATPase). Such ATPases are thought to be indirectly involved in active K(+) uptake, which can be facilitated by ATPase-dependent generation of an electrical driving force. Thus, K(+) effects on the chloroplast enzyme in vitro were found to be consistent with the hypothesized role of this envelope ATPase in facilitating active cation transport in vivo.