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. 1984 Oct;76(2):490–497. doi: 10.1104/pp.76.2.490

Anion-Sensitive, H+-Pumping ATPase of Oat Roots 1

Direct Effects of Cl−, NO3−, and a Disulfonic Stilbene

Kathleen A Churchill 1,2,3,2, Heven Sze 1,2,3
PMCID: PMC1064316  PMID: 16663870

Abstract

To understand the mechanism and molecular properties of the tonoplast-type H+-translocating ATPase, we have studied the effect of Cl−, NO3−, and 4,4′-diisothiocyano-2,2′-stilbene disulfonic acid (DIDS) on the activity of the electrogenic H+-ATPase associated with low-density microsomal vesicles from oat roots (Avena sativa cv Lang). The H+-pumping ATPase generates a membrane potential (Δψ) and a pH gradient (ΔpH) that make up two interconvertible components of the proton electrochemical gradient (Δ̄μh+). A permeant anion (e.g. Cl−), unlike an impermeant anion (e.g. iminodiacetate), dissipated the membrane potential ([14C]thiocyanate distribution) and stimulated formation of a pH gradient ([14C]methylamine distribution). However, Cl−-stimulated ATPase activity was about 75% caused by a direct stimulation of the ATPase by Cl− independent of the proton electrochemical gradient. Unlike the plasma membrane H+-ATPase, the Cl−-stimulated ATPase was inhibited by NO3− (a permeant anion) and by DIDS. In the absence of Cl−, NO3− decreased membrane potential formation and did not stimulate pH gradient formation. The inhibition by NO3− of Cl−-stimulated pH gradient formation and Cl−-stimulated ATPase activity was noncompetitive. In the absence of Cl−, DIDS inhibited the basal Mg,ATPase activity and membrane potential formation. DIDS also inhibited the Cl−-stimulated ATPase activity and pH gradient formation. Direct inhibition of the electrogenic H+-ATPase by NO3− or DIDS suggest that the vanadate-insensitive H+-pumping ATPase has anion-sensitive site(s) that regulate the catalytic and vectorial activity. Whether the anion-sensitive H+-ATPase has channels that conduct anions is yet to be established.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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