Cold inactivation of vacuolar proton-ATPases.

Y. Moriyama, N. Nelson

Research output: Contribution to journalArticle

155 Citations (Scopus)

Abstract

Incubation of the reconstituted H+-ATPase from chromaffin granules on ice resulted in inactivation of the proton-pumping and ATPase activities of the enzyme. Inactivation was dependent on the presence of Mg2+, Cl-, and ATP during the incubation at low temperature. Approximately 1 mM ATP, 1 mM Mg2+, and 200 mM Cl- were required for maximum inactivation. Incubation for about 10 min on ice was required to achieve 50% inactivation. A much smaller decline in activity was observed when the enzyme was incubated at room temperature with the same chemicals. Inactivation in the cold resulted in the release of five polypeptides from the membrane with apparent molecular masses of 72, 57, 41, 34, and 33 kDa on sodium dodecyl sulfate gels. Three of the polypeptides of 72, 57, and 34 kDa were identified as subunits of vacuolar H+-ATPases by antibody cross-reactivity. Similar results were obtained with several other vacuolar H+-ATPases including those from plant sources. It was concluded that the catalytic sector of the enzyme is released from the H+-ATPase complex by cold treatment, resulting in inactivation of the enzyme.

Original languageEnglish
Pages (from-to)3577-3582
Number of pages6
JournalJournal of Biological Chemistry
Volume264
Issue number6
Publication statusPublished - Feb 25 1989
Externally publishedYes

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Vacuolar Proton-Translocating ATPases
Adenosine Triphosphatases
Protons
Proton-Translocating ATPases
Ice
Enzymes
Adenosine Triphosphate
Chromaffin Granules
Peptides
Temperature
Molecular mass
Sodium Dodecyl Sulfate
Gels
Membranes
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Moriyama, Y., & Nelson, N. (1989). Cold inactivation of vacuolar proton-ATPases. Journal of Biological Chemistry, 264(6), 3577-3582.

Cold inactivation of vacuolar proton-ATPases. / Moriyama, Y.; Nelson, N.

In: Journal of Biological Chemistry, Vol. 264, No. 6, 25.02.1989, p. 3577-3582.

Research output: Contribution to journalArticle

Moriyama, Y & Nelson, N 1989, 'Cold inactivation of vacuolar proton-ATPases.', Journal of Biological Chemistry, vol. 264, no. 6, pp. 3577-3582.
Moriyama Y, Nelson N. Cold inactivation of vacuolar proton-ATPases. Journal of Biological Chemistry. 1989 Feb 25;264(6):3577-3582.
Moriyama, Y. ; Nelson, N. / Cold inactivation of vacuolar proton-ATPases. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 6. pp. 3577-3582.
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