Inhibition and Labeling of the Plant Plasma Membrane H-ATPase with N-Ethylmaleimide

Overview

Journal Plant Physiol

Specialty Physiology

Date 1987 Apr 1

PMID 16665375

Citations 6

Authors

D B Katz

M R Sussman

Affiliations

Soon will be listed here.

Abstract

H(+)-ATPase activity in plasma membranes isolated from Avena sativa root cells is inhibited by N-ethylmaleimide, a covalent modifier of protein sulfhydryl groups. The rate of inhibition is reduced by ADP, MgADP, and MgATP, but even at 40 millimolar ADP the enzyme is only partially protected against inactivation. When plasma membranes are treated wth N-[2-(3)H]ethylmaleimide and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, prominent radioactive bands appear at M(r)=100,000 and several other positions. However, only radioactivity in the M(r)=100,000 protein is reduced by the presence of MgADP. These results provide independent evidence that the M(r)=100,000 polypeptide which is observed in purified preparations of the enzyme is the catalytic subunit of the H(+)-ATPase. When tryptic peptides are produced from N-[2-(3)H]ethylmaleimide labeled M(r)=100,000 protein and separated by reverse phase high performance liquid chromatography, two radioactive peaks are observed for which N-[2-(3)H]ethylmaleimide incorporation is reduced in the presence of MgADP.

Citing Articles

Role of the Plasmalemma H-ATPase in Pseudomonas syringae-Induced K/H Exchange in Suspension-Cultured Tobacco Cells.

Atkinson M, Baker C Plant Physiol. 1989; 91(1):298-303.

PMID: 16667014 PMC: 1061990. DOI: 10.1104/pp.91.1.298.

Cytochemical localization of ATPase activity in oat roots localizes a plasma membrane-associated soluble phosphatase, not the proton pump.

Katz D, Sussman M, Mierzwa R, Evert R Plant Physiol. 1988; 86(3):841-7.

PMID: 16665998 PMC: 1054581. DOI: 10.1104/pp.86.3.841.

Isolation and sequence of tryptic peptides from the proton-pumping ATPase of the oat plasma membrane.

Schaller G, Sussman M Plant Physiol. 1988; 86(2):512-6.

PMID: 16665938 PMC: 1054515. DOI: 10.1104/pp.86.2.512.

A Sulfhydryl Reagent Modulates Systemic Signaling for Wound-Induced and Systemin-Induced Proteinase Inhibitor Synthesis.

Narvaez-Vasquez J, Orozco-Cardenas M, Ryan C Plant Physiol. 1994; 105(2):725-730.

PMID: 12232239 PMC: 159414. DOI: 10.1104/pp.105.2.725.

Effects of Prolonged Washing on Primary and Secondary Transport Processes at the Plasma Membrane in Red Beet Storage Tissue.

Marvier A, Williams L, Leigh R, Hall J Plant Physiol. 2002; 115(1):263-272.

PMID: 12223806 PMC: 158482. DOI: 10.1104/pp.115.1.263.

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