Bafilomycin inhibits proton flow through the H+ channel of vacuolar proton pumps

Bill P. Crider, Xiao Song Xie, Dennis K. Stone

Research output: Contribution to journalArticle

131 Citations (Scopus)

Abstract

Vacuolar-type proton-translocating ATPases are complex heterooligomers that are characterized by a specific inhibition by bafilomycin A1. These enzymes have a peripheral ATP hydrolytic domain as well as a transmembranous sector. The transmembranous sector has been isolated by glycerol gradient centrifugation, and this subcomplex is composed of polypeptides of 116, 39, and 17 kDa. Both this sector and native holoenzyme were reconstituted into potassium-loaded (150 mM KCl) liposomes prepared from pure lipids. When diluted into potassium-free buffer, a valinomycin-induced membrane potential did not drive proton uptake, as assessed by acridine orange quenching. In contrast, pretreatment of both the reconstituted proton pump and isolated transmembranous sector at pH 4.2 activated a latent proton conductance. Bafilomycin A1 (1 nM) inhibited ATP-energized proton pumping catalyzed by the proton pump, as well as membrane potential-driven proton flow through both the acid-activated proton pump and the isolated proton pore. Thus bafilomycin A1 inhibits vacuolar proton pumps by blocking proton conduction through the proton pore, which we term V(B).

Original languageEnglish (US)
Pages (from-to)17379-17381
Number of pages3
JournalJournal of Biological Chemistry
Volume269
Issue number26
StatePublished - Jul 1 1994

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Proton Pumps
Protons
Membrane Potentials
Potassium
Adenosine Triphosphate
Vacuolar Proton-Translocating ATPases
Membranes
Valinomycin
Holoenzymes
Acridine Orange
Proton-Translocating ATPases
Centrifugation
Liposomes
Glycerol
Quenching
Buffers
Lipids
Peptides
Acids
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Bafilomycin inhibits proton flow through the H+ channel of vacuolar proton pumps. / Crider, Bill P.; Xie, Xiao Song; Stone, Dennis K.

In: Journal of Biological Chemistry, Vol. 269, No. 26, 01.07.1994, p. 17379-17381.

Research output: Contribution to journalArticle

Crider, Bill P. ; Xie, Xiao Song ; Stone, Dennis K. / Bafilomycin inhibits proton flow through the H+ channel of vacuolar proton pumps. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 26. pp. 17379-17381.
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