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 Citations (Scopus)

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 - Nov 5 1993

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Clathrin-Coated Vesicles
Clathrin
Proton-Translocating ATPases
Adenosine Triphosphate
Proton Pumps
Ethylmaleimide
Molecular mass
Catalysis
Recombinant Proteins
Escherichia coli
Hydrolysis
Calcium

ASJC Scopus subject areas

  • Biochemistry

Cite this

Reconstitution of recombinant 40-kDa subunit of the clathrin-coated vesicle H+-ATPase. / Peng, Sheng Bin; Stone, Dennis K.; Xie, Xiao Song.

In: Journal of Biological Chemistry, Vol. 268, No. 31, 05.11.1993, p. 23519-23523.

Research output: Contribution to journalArticle

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