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. 1983 Mar;71(3):610–617. doi: 10.1104/pp.71.3.610

Anion-Sensitive, H+-Pumping ATPase in Membrane Vesicles from Oat Roots 1

Kathleen A Churchill 1,2,3, Heven Sze 1,2,3
PMCID: PMC1066086  PMID: 16662875

Abstract

H+-pumping ATPases were detected in microsomal vesicles of oat (Avena sativa L. var Lang) roots using [14C]methylamine distribution or quinacrine fluorescent quenching. Methylamine (MeA) accumulation into vesicles and quinacrine quench were specifically dependent on Mg,ATP. Both activities reflected formation of a proton gradient (ΔpH) (acid inside) as carbonyl cyanide m-chlorophenylhydrazone, nigericin (in the presence of K+), or gramicidin decreased MeA uptake or increased quinacrine fluorescence. The properties of H+ pumping as measured by MeA uptake were characterized. The Kmapp for ATP was about 0.1 millimolar. Mg,GTP and Mg, pyrophosphate were 19% and 30% as effective as Mg,ATP. MeA uptake was inhibited by N,N′-dicyclohexylcarbodiimide and was mostly insensitive to oligomycin, vanadate, or copper. ATP-dependent MeA was stimulated by anions with decreasing order of potency of Cl− > Br− > NO3− > SO42−, iminodiacetate, benzene sulfonate. Anion stimulation of H+ pumping was caused in part by the ability of permeant anions to dissipate the electrical potential and in part by a specific requirement of Cl− by a H+ -pumping ATPase. A pH gradient, probably caused by a Donnan potential, could be dissipated by K+ in the presence or absence of ATP. MeA uptake was enriched in vesicles of relatively low density and showed a parallel distribution with vanadate-insensitive ATPase activity on a continuous dextran gradient. ΔpH as measured by quinacrine quench was partially vanadate-sensitive. These results show that plant membranes have at least two types of H+ -pumping ATPases. One is vanadate-sensitive and probably enriched in the plasma membrane. One is vanadate-resistant, anion-sensitive and has many properties characteristic of a vacuolar ATPase. These results are consistent with the presence of electrogenic H+ pumps at the plasma membrane and tonoplast of higher plant cells.

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Selected References

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

  1. Cretin H. The proton gradient across the vacuo-lysosomal membrane of lutoids from the latex of Hevea brasiliensis. I. Further evidence for a proton-translocating ATPase on the vacuo-lysosomal membrane of intact lutoids. J Membr Biol. 1982;65(3):175–184. doi: 10.1007/BF01869961. [DOI] [PubMed] [Google Scholar]
  2. Dupont F. M., Bennett A. B., Spanswick R. M. Localization of a proton-translocating ATPase on sucrose gradients. Plant Physiol. 1982 Oct;70(4):1115–1119. doi: 10.1104/pp.70.4.1115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Kakinuma Y., Ohsumi Y., Anraku Y. Properties of H+-translocating adenosine triphosphatase in vacuolar membranes of SAccharomyces cerevisiae. J Biol Chem. 1981 Nov 10;256(21):10859–10863. [PubMed] [Google Scholar]
  4. Kleiner D. The transport of NH3 and NH4+ across biological membranes. Biochim Biophys Acta. 1981 Nov 9;639(1):41–52. doi: 10.1016/0304-4173(81)90004-5. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Lee H. C., Forte J. G. A study of H+ transport in gastric microsomal vesicles using fluorescent probes. Biochim Biophys Acta. 1978 Apr 4;508(2):339–356. doi: 10.1016/0005-2736(78)90336-x. [DOI] [PubMed] [Google Scholar]
  7. Leonard R. T., Hodges T. K. Characterization of Plasma Membrane-associated Adenosine Triphosphase Activity of Oat Roots. Plant Physiol. 1973 Jul;52(1):6–12. doi: 10.1104/pp.52.1.6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lilley R. M. Isolation of Functionally Intact Rhodoplasts from Griffithsia monilis (Ceramiaceae, Rhodophyta). Plant Physiol. 1981 Jan;67(1):5–8. doi: 10.1104/pp.67.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Linnett P. E., Beechey R. B. Inhibitors of the ATP synthethase system. Methods Enzymol. 1979;55:472–518. doi: 10.1016/0076-6879(79)55061-7. [DOI] [PubMed] [Google Scholar]
  11. Marin B., Blasco F. Further evidence for the proton pumping work of tonoplast ATPase from Hevea latex vacuome. Biochem Biophys Res Commun. 1982 Mar 15;105(1):354–361. doi: 10.1016/s0006-291x(82)80052-1. [DOI] [PubMed] [Google Scholar]
  12. Marin B., Marin-Lanza M., Komor E. The protonmotive potential difference across the vacuo-lysosomal membrane of Hevea brasiliensis (rubber tree) and its modification by a membrane-bound adenosine triphosphatase. Biochem J. 1981 Aug 15;198(2):365–372. doi: 10.1042/bj1980365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Raven J. A., Farquhar G. D. Methylammonium Transport in Phaseolus vulgaris Leaf Slices. Plant Physiol. 1981 Apr;67(4):859–863. doi: 10.1104/pp.67.4.859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rottenberg H., Grunwald T., Avron M. Determination of pH in chloroplasts. I. Distribution of ( 14 C) methylamine. Eur J Biochem. 1972 Jan 31;25(1):54–63. doi: 10.1111/j.1432-1033.1972.tb01666.x. [DOI] [PubMed] [Google Scholar]
  15. Rottenberg H. The measurement of membrane potential and deltapH in cells, organelles, and vesicles. Methods Enzymol. 1979;55:547–569. doi: 10.1016/0076-6879(79)55066-6. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Scarborough G. A. Proton translocation catalyzed by the electrogenic ATPase in the plasma membrane of Neurospora. Biochemistry. 1980 Jun 24;19(13):2925–2931. doi: 10.1021/bi00554a017. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. 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]
  21. 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]

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