New evidence for the dimeric nature of NADH:Q oxidoreductase in bovine-heart submitochondrial particles.
The initial velocity of NADH oxidation by bovine-heart submitochondrial particles was measured at pH 8.0 after pretreatment of these particles with different amounts of the inhibitor piericidine A together with 0.035 mM NADH. The amount of piericidine A required to fully inhibit the NADH oxidation activity extrapolated to exactly 1.0 per Fe-S cluster 2 of NADH:Q oxidoreductase. When no reducing equivalents from NADH were present during the pretreatment, this ratio was 1.2. The difference is explained by assuming that NADH:Q oxidoreductase binds piericidine A more effectively in the reduced state than in the oxidized state. It was also found that after Q10-extraction and reincorporation of submitochondrial particles, the amount of piericidine A required to fully inhibit the NADH oxidation activity of the particles increased with the amount of Q10 present during reincorporation. This is explained by assuming that binding of piericidine A, to the inhibitory site of NADH:Q oxidoreductase requires Q10. When 0.035 mM NADPH instead of NADH was present during the pretreatment of submitochondrial particles with piericidine A, the amount of inhibitor per cluster 2 required to fully inhibit the initial NADH-oxidation activity extrapolated to 0.5. This result strongly suggests that NADH:Q oxidoreductase is a functional dimer.
van Belzen
R
R
E.C. Slater Institute for Biochemical Research, University of Amsterdam, The Netherlands.
van Gaalen
M C
MC
Cuypers
P A
PA
Albracht
S P
SP
D000818:Animals; D002417:Cattle; D004579:Electron Transport; D006863:Hydrogen-Ion Concentration; D008929:Mitochondria, Heart / Q000201:enzymology*; D015394:Molecular Structure; D016660:NAD(P)H Dehydrogenase (Quinone); D009249:NADP / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D011808:Quinone Reductases / Q000037:antagonists & inhibitors*
D011725:Pyridines; D009249:NADP; C100213:piericidin A; D016660:NAD(P)H Dehydrogenase (Quinone); D011808:Quinone Reductases