Measurement of matrix enzyme activity in isolated mitochondria made permeable with toluene

M. Abdul Matlib, W. Allen Shannon, P. Srere

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Mitochondria isolated from rat liver and heart were made permeable to normally nonpentrating substrates and cofactors by treatment with toluene. The optimal conditions for preparing stable, permeable mitochondria were 2% toluene for 2 min at 4 °C in a buffered, isotonic medium containing 8.5% polyethylene glycol (Mr 6000-7500). Without polyethylene glycol, the toluene-treated mitochondria were unstable and released their matrix enzymes. The treated mitochondria were particularly unstable in dilute suspension under normal assay conditions of their enzyme activities. The levels of matrix enzyme activities unmasked by toluene treatment of mitochondria were very close to those of sonicated mitochondria under identical assay conditions. Mitochondria made permeable with toluene lost only small amounts of their protein and retained a major fraction of the nucleotides and coenzymes. Electron microscopic examination of toluenetreated mitochondria indicated that they were relatively intact with swollen and vesiculated cristae membranes. Such preparations will allow the study of mitochondrial enzymes at approximate in vivo concentrations.

Original languageEnglish
Pages (from-to)396-407
Number of pages12
JournalArchives of Biochemistry and Biophysics
Volume178
Issue number2
DOIs
Publication statusPublished - Jan 30 1977

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Mitochondria
Toluene
Enzyme activity
Enzymes
Assays
Coenzymes
Liver
Rats
Suspensions
Microscopic examination
Nucleotides
Electrons
Membranes
Substrates

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Measurement of matrix enzyme activity in isolated mitochondria made permeable with toluene. / Matlib, M. Abdul; Shannon, W. Allen; Srere, P.

In: Archives of Biochemistry and Biophysics, Vol. 178, No. 2, 30.01.1977, p. 396-407.

Research output: Contribution to journalArticle

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