An ATP-driven proton pump in clathrin-coated vesicles.

D. K. Stone, X. S. Xie, E. Racker

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Abstract

Clathrin containing coated vesicles prepared from bovine brain catalyzed ATP-driven proton translocation and a 32Pi-ATP exchange reaction. Both activities were measured in the presence of 5 micrograms of oligomycin/mg of protein which completely inhibited these reactions catalyzed by submitochondrial particles. Analyses performed during the purification procedure demonstrated that the oligomycin-resistant pump was concentrated and highly purified in the fractions containing coated vesicles. Moreover, vesicles precipitated by either monoclonal or polyclonal antibodies against clathrin contained the H+ pump activity. Dicyclohexylcarbodiimide (0.5 mM) and N-ethylmaleimide (1 mM) added to the assay mixture inhibited the pump completely, whereas neither vanadate, sodium azide, efrapeptin, or mitochondrial ATPase inhibitor had an effect.

Original languageEnglish (US)
Pages (from-to)4059-4062
Number of pages4
JournalJournal of Biological Chemistry
Volume258
Issue number7
Publication statusPublished - Apr 10 1983

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ASJC Scopus subject areas

  • Biochemistry

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