Energy-linked nicotinamide nucleotide transhydrogenase: hydrodynamic properties and active form of purified and membrane-bound mitochondrial transhydrogenase from beef heart.
The mitochondrial nicotinamide nucleotide transhydrogenase from beef heart was investigated with respect to minimal assembly of the purified enzyme and of the enzyme in the mitochondrial inner membrane. Studies of the hydrodynamic properties of the purified enzyme in the presence of 0.3% Triton X-100 allowed determination of the Stokes radius, sedimentation constant, partial specific volume, frictional ratio, and molecular weight. Under these conditions transhydrogenase existed as an inactive monomer, suggesting that monomerization may be accompanied by inactivation. Radiation inactivation was used to determine the functional molecular size of purified detergent-dispersed transhydrogenase and transhydrogenase in beef heart submitochondrial particles. Under these conditions the catalytic activity of both the purified and the membrane-bound enzyme was found to be catalyzed by a dimeric form of the enzyme. These results suggest for the first time that the minimal functional assembly of detergent-dispersed as well as membrane-bound transhydrogenase is a dimer, which is not functionally associated with, for example, complex I or ATPase. In addition, the results are consistent with the possibility that the two subunits of transhydrogenase are catalytically active in an alternating fashion according to a previously proposed half-of-the-sites reactivity model.
Department of Biochemistry, University of Stockholm, Sweden.
B
B
Persson
G
G
Ahnström
J
J
Rydström
D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D008929:Mitochondria, Heart / Q000201:enzymology*; D008970:Molecular Weight; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009250:NADP Transhydrogenases / Q000378:metabolism*; D017830:Octoxynol; D011092:Polyethylene Glycols; D012995:Solubility; D013367:Submitochondrial Particles / Q000201:enzymology
D011092:Polyethylene Glycols; D017830:Octoxynol; D009247:NADH, NADPH Oxidoreductases; D009250:NADP Transhydrogenases