Proton ATPase of rat liver mitochondria: A rapid procedure for purification of a stable, reconstitutively active F1 preparation using a modified chloroform method

Noreen Williams, Mario L Amzel, Peter L Pedersen

Research output: Contribution to journalArticle

Abstract

A method is described for the purification of rat liver F1-ATPase by a modification of the chloroform extraction procedure originally described by Beechey et al. (Biochem. J. (1975) 148, 533). Purified liver membrane vesicles are extracted with chloroform in the presence of ATP and EDTA. The procedure yields pure F1 in only 2-3 h without the necessity of ion-exchange chromatography. The enzyme exhibits the α, β, γ, δ, and ε{lunate} bands characteristic of F1-ATPase. It has a high ATPase specific activity, and is reconstitutively active, catalyzing high rates of ATP synthesis. Significantly, it can be readily crystallized. If desired, the enzyme can be passed over a gel filtration column to place it in a stabilizing phosphate-EDTA buffer, lyophilized and stored indefinitely at -20°C.

Original languageEnglish (US)
Pages (from-to)581-588
Number of pages8
JournalAnalytical Biochemistry
Volume140
Issue number2
DOIs
StatePublished - Aug 1 1984

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Mitochondria
Proton-Translocating ATPases
Liver Mitochondrion
Chloroform
Edetic Acid
Liver
Purification
Adenosine Triphosphatases
Protons
Rats
Adenosine Triphosphate
Enzymes
Chromatography
Ion exchange
Buffers
Gels
Phosphates
Ion Exchange Chromatography
Membranes
Gel Chromatography

Keywords

  • F-ATPase, ATP synthase, ATPase, FF-ATPase, mitochondrial ATPase
  • proton ATPase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Proton ATPase of rat liver mitochondria : A rapid procedure for purification of a stable, reconstitutively active F1 preparation using a modified chloroform method. / Williams, Noreen; Amzel, Mario L; Pedersen, Peter L.

In: Analytical Biochemistry, Vol. 140, No. 2, 01.08.1984, p. 581-588.

Research output: Contribution to journalArticle

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