Purification of the Proton-Translocating ATPase from Rat Liver Mitochondria Using the Detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate

Maureen W. McEnery, Peter L. Pedersen

Research output: Contribution to journalArticlepeer-review

Abstract

This chapter describes the procedure for the purification of the proton-translocating ATPase from rat liver mitochondria using the detergent 3-[(3-Cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS). The important feature of the procedure that follows for the purification of the F0F1-ATPase from rat liver mitochondria that distinguishes it from other procedures of purification of F0F1ATPase complexes is the use of the detergent CHAPS that is developed as an alternative to existing detergents. CHAPS appears to be the detergent of choice for the solubilization of the membrane-bound F0F1ATPase complex. A change in aggregation of the complex forms the basis for the purification of the F0F1ATPase complex. This modification simplifies the isolation procedure and eliminates the problems that occur when trying to control for the presence of sucrose in the aliquots assayed for protein concentration via the Bradford protein assay. The purified and reconstituted complex catalyzes ATP-Pi exchange and ATP-dependent proton translocation.

Original languageEnglish (US)
Pages (from-to)470-477
Number of pages8
JournalMethods in enzymology
Volume126
Issue numberC
DOIs
StatePublished - Jan 1 1986

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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