Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 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.

Full text

PDF
490

Selected References

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

  1. Bennett A. B., O'neill S. D., Spanswick R. M. H-ATPase Activity from Storage Tissue of Beta vulgaris: I. Identification and Characterization of an Anion-Sensitive H-ATPase. Plant Physiol. 1984 Mar;74(3):538–544. doi: 10.1104/pp.74.3.538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Briskin D. P., Leonard R. T. Partial characterization of a phosphorylated intermediate associated with the plasma membrane ATPase of corn roots. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6922–6926. doi: 10.1073/pnas.79.22.6922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cabantchik Z. I., Knauf P. A., Rothstein A. The anion transport system of the red blood cell. The role of membrane protein evaluated by the use of 'probes'. Biochim Biophys Acta. 1978 Sep 29;515(3):239–302. doi: 10.1016/0304-4157(78)90016-3. [DOI] [PubMed] [Google Scholar]
  4. Cantley L. C., Jr, Cantley L. G., Josephson L. A characterization of vanadate interactions with the (Na,K)-ATPase. Mechanistic and regulatory implications. J Biol Chem. 1978 Oct 25;253(20):7361–7368. [PubMed] [Google Scholar]
  5. Churchill K. A., Holaway B., Sze H. Separation of two types of electrogenic h-pumping ATPases from oat roots. Plant Physiol. 1983 Dec;73(4):921–928. doi: 10.1104/pp.73.4.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Churchill K. A., Sze H. Anion-sensitive, h-pumping ATPase in membrane vesicles from oat roots. Plant Physiol. 1983 Mar;71(3):610–617. doi: 10.1104/pp.71.3.610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dupont F. M., Giorgi D. L., Spanswick R. M. Characterization of a proton-translocating ATPase in microsomal vesicles from corn roots. Plant Physiol. 1982 Dec;70(6):1694–1699. doi: 10.1104/pp.70.6.1694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gallagher S. R., Leonard R. T. Effect of vanadate, molybdate, and azide on membrane-associated ATPase and soluble phosphatase activities of corn roots. Plant Physiol. 1982 Nov;70(5):1335–1340. doi: 10.1104/pp.70.5.1335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hatefi Y., Hanstein W. G. Destabilization of membranes with chaotropic ions. Methods Enzymol. 1974;31:770–790. doi: 10.1016/0076-6879(74)31080-4. [DOI] [PubMed] [Google Scholar]
  10. Hodges T. K., Leonard R. T. Purification of a plasma membrane-bound adenosine triphosphatase from plant roots. Methods Enzymol. 1974;32:392–406. doi: 10.1016/0076-6879(74)32039-3. [DOI] [PubMed] [Google Scholar]
  11. Keifer D. W., Franceschi V. R., Lucas W. J. Plasmalemma Chloride Transport in Chara corallina: Inhibition by 4,4'-Diisothiocyano-2,2'-Disulfonic Acid Stilbene. Plant Physiol. 1982 Nov;70(5):1327–1334. doi: 10.1104/pp.70.5.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Lin W. Inhibition of anion transport in corn root protoplasts. Plant Physiol. 1981 Aug;68(2):435–438. doi: 10.1104/pp.68.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. MITCHELL P. Coupling of phosphorylation to electron and hydrogen transfer by a chemi-osmotic type of mechanism. Nature. 1961 Jul 8;191:144–148. doi: 10.1038/191144a0. [DOI] [PubMed] [Google Scholar]
  15. Mandala S., Mettler I. J., Taiz L. Localization of the proton pump of corn coleoptile microsomal membranes by density gradient centrifugation. Plant Physiol. 1982 Dec;70(6):1743–1747. doi: 10.1104/pp.70.6.1743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mettler I. J., Mandala S., Taiz L. Characterization of in vitro proton pumping by microsomal vesicles isolated from corn coleoptiles. Plant Physiol. 1982 Dec;70(6):1738–1742. doi: 10.1104/pp.70.6.1738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. O'neill S. D., Bennett A. B., Spanswick R. M. Characterization of a NO(3)-Sensitive H-ATPase from Corn Roots. Plant Physiol. 1983 Jul;72(3):837–846. doi: 10.1104/pp.72.3.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pazoles C. J., Creutz C. E., Ramu A., Pollard H. B. Permeant anion activation of MgATPase activity in chromaffin granules. Evidence for direct coupling of proton and anion transport. J Biol Chem. 1980 Aug 25;255(16):7863–7869. [PubMed] [Google Scholar]
  19. Perlin D. S., Kasamo K., Brooker R. J., Slayman C. W. Electrogenic H+ translocation by the plasma membrane ATPase of Neurospora. Studies on plasma membrane vesicles and reconstituted enzyme. J Biol Chem. 1984 Jun 25;259(12):7884–7892. [PubMed] [Google Scholar]
  20. Poole R. J., Briskin D. P., Krátký Z., Johnstone R. M. Density gradient localization of plasma membrane and tonoplast from storage tissue of growing and dormant red beet : characterization of proton-transport and ATPase in tonoplast vesicles. Plant Physiol. 1984 Mar;74(3):549–556. doi: 10.1104/pp.74.3.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rungie J. M., Wiskich J. T. Salt-stimulated Adenosine Triphosphatase from Smooth Microsomes of Turnip. Plant Physiol. 1973 Jun;51(6):1064–1068. doi: 10.1104/pp.51.6.1064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stout R. G., Cleland R. E. Evidence for a Cl-Stimulated MgATPase Proton Pump in Oat Root Membranes. Plant Physiol. 1982 Apr;69(4):798–803. doi: 10.1104/pp.69.4.798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sze H. Characterization of nigericin-stimulated ATPase from sealed microsomal vesicles of tobacco callus. Plant Physiol. 1982 Aug;70(2):498–505. doi: 10.1104/pp.70.2.498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sze H., Churchill K. A. Mg/KCl-ATPase of plant plasma membranes is an electrogenic pump. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5578–5582. doi: 10.1073/pnas.78.9.5578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sze H. Nigericin-stimulated ATPase activity in microsomal vesicles of tobacco callus. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5904–5908. doi: 10.1073/pnas.77.10.5904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vara F., Serrano R. Partial purification and properties of the proton-translocating ATPase of plant plasma membranes. J Biol Chem. 1982 Nov 10;257(21):12826–12830. [PubMed] [Google Scholar]
  27. Xie X. S., Stone D. K., Racker E. Determinants of clathrin-coated vesicle acidification. J Biol Chem. 1983 Dec 25;258(24):14834–14838. [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES