title: ATP synthesis catalyzed by a V-ATPase: an alternative pathway for energy conservation operating in plant vacuoles?
abstract: The electrochemical H(+) gradient generated in tonoplast vesicles isolated from maize seeds was found to be able to drive the reversal of the catalytic cycle of both vacuolar H(+)-pumps (Façanha and de Meis, 1998). Here we describe the reversibility of the vacuolar V-type H(+)-ATPase (V-ATPase) even in the absence of the H(+) gradient in a water-Me2SO co-solvent mixture, resulting in net synthesis of [γ-(32)P]ATP from [(32)P]Pi and ADP. The water-Me2SO (5 to 20 %) media promoted inhibition of both PPi hydrolysis and synthesis reactions whereas it slightly affected the ATP hydrolysis and clearly stimulated the ATP synthesis, which was unaffected by uncoupling agents (FCCP, Triton X-100 or NH4 (+)). This effect of Me2SO on the ATP⇔(32)P exchange reaction seems to be related to a decrease of the apparent K m of the V-ATPase for Pi. The results are in accordance to the concept that the energetics of ATP synthesis catalysis depends on the solvation energies interacting in the enzyme microenvironment. A possible physiological significance of this phenomenon for the metabolism of desiccation-tolerant plant cells is discussed.
authors.affiliation: Laboratório de Biologia Celular & Tecidual, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Av. Alberto Lamego, 2000, Pq. California, Campos dos Goytacazes, RJ 28013-602 Brazil.
authors.forename: Arnoldo Rocha
authors.initials: AR
authors.lastname: Façanha
authors.forename: Anna Lvovna
authors.initials: AL
authors.lastname: Okorokova-Façanha
keywords: DMSO; V1V0-ATPase; bind energy; corn seeds; membrane bound H+-pyrophosphatase; proton gradient; proton pumps