Reconstitution of recombinant 40-kDa subunit of the clathrin-coated vesicle H+-ATPase

Sheng Bin Peng, Dennis K. Stone, Xiao Song Xie

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

We have proposed a model of the ATP hydrolytic sector of the clathrin- coated vesicle H+-ATPase wherein significant catalysis requires four subunits of molecular masses of 70, 58, 40, and 33 kDa (Xie, X.-S., and Stone, D. K. (1988) J. Biol. Chem. 263, 9859-9867). We have cloned and expressed the 40-kDa component in Escherichia coli and have purified the recombinant protein to homogeneity. This subunit lacks ATP hydrolytic capacity, but when reconstituted to a 40 kDa-depleted hydrolytic sector, there is a greater than 20-fold increase in calcium-activated, N- ethylmaleimide-sensitive ATP hydrolysis, indicating that this subunit is required for vacuolar-type proton pump function.

Original languageEnglish (US)
Pages (from-to)23519-23523
Number of pages5
JournalJournal of Biological Chemistry
Volume268
Issue number31
StatePublished - Jan 1 1993

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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