title: Fusicoccin Activates the Plasma Membrane H+-ATPase by a Mechanism Involving the C-Terminal Inhibitory Domain.
abstract: Plasma membrane vesicles isolated from spinach leaves incubated with the fungal toxin fusicoccin showed a twofold increase in ATP hydrolytic activity and a threefold increase in H+ pumping compared to controls. This increase in H+-ATPase activity was largely completed within 4 min of incubation and was not due to de novo synthesis of H+-ATPase as demonstrated by immunoblotting. Incubation with fusicoccin also resulted in a decrease in the apparent Km for ATP of the H+-ATPase from 0.22 to 0.10 mM. The fusicoccin-mediated activation of H+-ATPase activity and the accompanying decrease in the Km for ATP are changes very similar to those observed upon trypsin activation of the H+-ATPase, where an autoinhibitory domain in the C-terminal region of the H+-ATPase is removed. Thus, trypsin treatment of plasma membrane vesicles from control leaves gave a twofold increase in ATP hydrolytic activity and a threefold increase in H+ pumping, as well as a decrease in the apparent Km for ATP of the H+-ATPase from 0.22 to 0.10 mM. Trypsin treatment of plasma membranes from fusicoccin-incubated leaves did not further enhance the H+-ATPase activity, however, and neither was the Km for ATP further decreased. That trypsin really removed a small segment from the fusicoccin-activated H+-ATPase was confirmed by immunoblotting, which showed the appearance of a 90-kD band in addition to the native 100-kD H+-ATPase band upon trypsin treatment. Taken together, our data suggest that in vivo activation of the H+-ATPase by fusicoccin proceeds by a mechanism involving a displacement of the C-terminal inhibitory domain.
authors.affiliation: Department of Plant Biochemistry, University of Lund, P.O. Box 7007, S-220 07 Lund, Sweden.
authors.forename: F.
authors.initials: F
authors.lastname: Johansson
authors.forename: M.
authors.initials: M
authors.lastname: Sommarin
authors.forename: C.
authors.initials: C
authors.lastname: Larsson