Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1982 Nov;70(5):1459–1464. doi: 10.1104/pp.70.5.1459

Phosphorylation of the Adenosine Triphosphatase in a Deoxycholate-Treated Plasma Membrane Fraction from Corn Roots 1

Donald P Briskin 1,2, Robert T Leonard 1
PMCID: PMC1065906  PMID: 16662698

Abstract

The ATP phosphohydrolase (ATPase) activity of a corn (Zea mays L., WF9 × Mo17) root plasma membrane fraction was enriched almost 2-fold by selective extraction with 0.1% (w/v) deoxycholate. The detergent treatment solubilized about 30% of the total membrane protein and some ATP hydrolyzing activity that was not K+-stimulated, but the major portion of the ATPase activity could be pelleted with membranes. The properties of the ATPase associated with the detergent-extracted plasma membrane fraction were similar to those for the ATPase of the untreated plasma membrane fraction with respect to substrate specificity, pH optimum, kinetics with MgATP, ion stimulation, and inhibitor sensitivity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed only minor differences in protein composition resulting from the detergent treatment.

The plasma membrane fraction from corn roots contained an endogenous protein kinase activity. This was shown by the time course of phosphate incorporation and by the labeling of a number of protein bands on SDS-polyacrylamide gel electrophoresis. The deoxycholate treatment removed measurable protein kinase activity and allowed the demonstration of a rapidly turning over covalent phosphorylated intermediate associated with the detergent-extracted plasma membrane fraction. The phosphorylated intermediate was present as a 100,000 dalton polypeptide and may represent the catalytic subunit of the plasma membrane K+-ATPase.

Full text

PDF
1459

Images in this article

1463Fig. 7
on p.1463

Selected References

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

  1. Amory A., Foury F., Goffeau A. The purified plasma membrane ATPase of the yeast Schizosaccharomyces pombe forms a phosphorylated intermediate. J Biol Chem. 1980 Oct 10;255(19):9353–9357. [PubMed] [Google Scholar]
  2. Briskin D. P., Leonard R. T. Isolation of tonoplast vesicles from tobacco protoplasts. Plant Physiol. 1980 Oct;66(4):684–687. doi: 10.1104/pp.66.4.684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Dame J. B., Scarborough G. A. Identification of the hydrolytic moiety of the Neurospora plasma membrane H+-ATPase and demonstration of a phosphoryl-enzyme intermediate in its catalytic mechanism. Biochemistry. 1980 Jun 24;19(13):2931–2937. doi: 10.1021/bi00554a018. [DOI] [PubMed] [Google Scholar]
  5. Dupont F. M., Burke L. L., Spanswick R. M. Characterization of a partially purified adenosine triphosphatase from a corn root plasma membrane fraction. Plant Physiol. 1981 Jan;67(1):59–63. doi: 10.1104/pp.67.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dupont F. M., Leonard R. T. Solubilization and partial purification of the adenosine triphosphatase from a corn root plasma membrane fraction. Plant Physiol. 1980 May;65(5):931–938. doi: 10.1104/pp.65.5.931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fairbanks G., Avruch J. Four gel systems for electrophoretic fractionation of membrane proteins using ionic detergents. J Supramol Struct. 1972;1(1):66–75. doi: 10.1002/jss.400010110. [DOI] [PubMed] [Google Scholar]
  8. Gunn R. B. Co- and counter-transport mechanisms in cell membranes. Annu Rev Physiol. 1980;42:249–259. doi: 10.1146/annurev.ph.42.030180.001341. [DOI] [PubMed] [Google Scholar]
  9. Hobbs A. S., Albers R. W. The structure of proteins involved in active membrane transport. Annu Rev Biophys Bioeng. 1980;9:259–291. doi: 10.1146/annurev.bb.09.060180.001355. [DOI] [PubMed] [Google Scholar]
  10. Knowles J. R. Enzyme-catalyzed phosphoryl transfer reactions. Annu Rev Biochem. 1980;49:877–919. doi: 10.1146/annurev.bi.49.070180.004305. [DOI] [PubMed] [Google Scholar]
  11. Kyte J. Purification of the sodium- and potassium-dependent adenosine triphosphatase from canine renal medulla. J Biol Chem. 1971 Jul 10;246(13):4157–4165. [PubMed] [Google Scholar]
  12. 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]
  13. Leonard R. T., Hotchkiss C. W. Cation-stimulated Adenosine Triphosphatase Activity and Cation Transport in Corn Roots. Plant Physiol. 1976 Sep;58(3):331–335. doi: 10.1104/pp.58.3.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Leonard R. T., Vanderwoude W. J. Isolation of plasma membranes from corn roots by sucrose density gradient centrifugation: an anomalous effect of ficoll. Plant Physiol. 1976 Jan;57(1):105–114. doi: 10.1104/pp.57.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lin P. P., Key J. L. Histone Kinase from Soybean Hypocotyls: PURIFICATION, PROPERTIES, AND SUBSTRATE SPECIFICITIES. Plant Physiol. 1980 Sep;66(3):360–367. doi: 10.1104/pp.66.3.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Malpartida F., Serrano R. Phosphorylated intermediate of the ATPase from the plasma membrane of yeast. Eur J Biochem. 1981 May 15;116(2):413–417. doi: 10.1111/j.1432-1033.1981.tb05350.x. [DOI] [PubMed] [Google Scholar]
  17. Meissner G., Conner G. E., Fleischer S. Isolation of sarcoplasmic reticulum by zonal centrifugation and purification of Ca 2+ -pump and Ca 2+ -binding proteins. Biochim Biophys Acta. 1973 Mar 16;298(2):246–269. doi: 10.1016/0005-2736(73)90355-6. [DOI] [PubMed] [Google Scholar]
  18. POST R. L., SEN A. K., ROSENTHAL A. S. A PHOSPHORYLATED INTERMEDIATE IN ADENOSINE TRIPHOSPHATE-DEPENDENT SODIUM AND POTASSIUM TRANSPORT ACROSS KIDNEY MEMBRANES. J Biol Chem. 1965 Mar;240:1437–1445. [PubMed] [Google Scholar]
  19. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  20. Peterson G. L. A simplified method for analysis of inorganic phosphate in the presence of interfering substances. Anal Biochem. 1978 Jan;84(1):164–172. doi: 10.1016/0003-2697(78)90495-5. [DOI] [PubMed] [Google Scholar]
  21. Sachs G., Berglindh T., Rabon E., Wallmark B., Barcellona M. L., Stewart H. B., Saccomani G. The interaction of K+ with gastric parietal cells and gastric ATPase. Ann N Y Acad Sci. 1980;358:118–137. doi: 10.1111/j.1749-6632.1980.tb15391.x. [DOI] [PubMed] [Google Scholar]
  22. Sarkadi B. Active calcium transport in human red cells. Biochim Biophys Acta. 1980 Sep 30;604(2):159–190. doi: 10.1016/0005-2736(80)90573-8. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. de Meis L., Vianna A. L. Energy interconversion by the Ca2+-dependent ATPase of the sarcoplasmic reticulum. Annu Rev Biochem. 1979;48:275–292. doi: 10.1146/annurev.bi.48.070179.001423. [DOI] [PubMed] [Google Scholar]

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

RESOURCES