The isolation of a basic coupling factor from preparations of factor F3.

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The isolation of a basic coupling factor from preparations of factor F3.

Publication Date
Sep 01, 1969
  • Biology
  • Ecology


Biochem. J. (1969) 114, 440 Printed in Great Britain The Isolation of a Basic Coupling Factor from Preparations of Factor F3 By R. B. 13EECHEY, I. G. K'NIGHT, C. R. LINDoP and K. J. CATTELL Shell Research Ltd., IVoodlstocke Agricultural Research Centre, Sittinybourne, Kent (Received 25 June 1969) A large numbiller of mitochondrial protein fractions that increase the efficiency of energy conservation in depleted mitochondrial-membrane preparations have been described over the past decade (Vallejos, Van den Berg & Slater, 1968). Precise comparison of the properties of those coupling factors that do not have ATPase* activity is very difficult. How- ever, there are sufficient similarities both in the methods of extraction of the coupling factors from mitochondria and in the funetional properties of these heterogeneous preparations to suggest that they may contain a common active component, which is positively charged in the pH range 7-8. We therefore decided to examine various pre- parations of coupling factors for the presence of a basic fraction that, in the presence of coupling factor F1 (mitochondrial ATPase), w-vould stimulate in A-particles (Lam, Warshaw & Sanadi, 1967) the ATP-driven reduction of NAD+ by suceinate and both the aerobic and ATP-driven energy-linked nicotinamide nucleotide transhydrogenase reactions and also confer oligomycin-sensitivity on factor F1 ATPase activity in the presence of TUA-particles (Bulos & Racker, 1968). The basicity of the protein fraction was established by its ability to adhere to CM-cellulose and the subsequent elution with 200mM-KCl-20mM-tris sulphate buffer, pH 8. These are among the properties of the oligo- mycin-sensitivity-conferring protein (MacLennan & Tzagoloff, 1968), but we believe that a title describ- ing a physical property rather than a putative function is more apt. We have termed our extract the basic coupling factor. The results presented here show the isolation of basic coupling factor from preparations of factor F3 (Fessenden & Racker

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