Isolation and comparative studies of mitochondrial F1-ATPase from Morris hepatoma and rat liver

Vitaly L. Spitsberg, H. P. Morris, Samuel H P Chan

Research output: Contribution to journalArticle

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Abstract

A simple method of isolating mitochondrial ATPase from rat liver and Morris hepatoma cell lines by chloroform extraction and chromatography on DEAE-Sephadex is described. This method is suitable even when small amounts of starting material with relatively low specific ATPase activity (in the case of hepatoma mitochondria and submitochondrial particles) are available. The isolated enzyme from both rat liver and hepatomas had a high specific activity, was similarly activated by bicarbonate and 2,4-dinitrophenol, and had a typical five-band pattern in sodium dodecyl sulfate electrophoresis. Prior to DEAE-Sephadex chromatography, an additional protein band which migrates between the δ and ε{lunate} subunits in the tumor F1-ATPase preparation was observed. The purified enzymes were cold labile and restored oxidative phosphorylation function of F1-ATPase depleted submitochondrial particles prepared from rat liver. The ATPase activity of the isolated enzymes was inhibited by mitochondrial ATPase inhibitor protein. The apparent stoichiometry of the inhibitor protein to the purified ATPase was extrapolated to be 2:1.

Original languageEnglish (US)
Pages (from-to)136-144
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume195
Issue number1
DOIs
StatePublished - 1979

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Experimental Liver Neoplasms
Proton-Translocating ATPases
Liver
Adenosine Triphosphatases
Rats
Submitochondrial Particles
DEAE-Dextran
Chromatography
Hepatocellular Carcinoma
Enzymes
2,4-Dinitrophenol
Proteins
Mitochondria
Oxidative Phosphorylation
Bicarbonates
Chloroform
Electrophoresis
Sodium Dodecyl Sulfate
Stoichiometry
Tumors

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Isolation and comparative studies of mitochondrial F1-ATPase from Morris hepatoma and rat liver. / Spitsberg, Vitaly L.; Morris, H. P.; Chan, Samuel H P.

In: Archives of Biochemistry and Biophysics, Vol. 195, No. 1, 1979, p. 136-144.

Research output: Contribution to journalArticle

Spitsberg, Vitaly L. ; Morris, H. P. ; Chan, Samuel H P. / Isolation and comparative studies of mitochondrial F1-ATPase from Morris hepatoma and rat liver. In: Archives of Biochemistry and Biophysics. 1979 ; Vol. 195, No. 1. pp. 136-144.
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