Protonophoric activity of NADH coenzyme Q reductase and ATP synthase in coupled submitochondrial particles from horse platelets.
A method to prepare coupled submitochondrial particles from horse platelets is described. The method allowed us to study the protonophoric activities of both complex I and complex V following the fluorescence quenching of the monoamine 9-amino-6-chloro-2 methoxyacridine (ACMA), a probe highly sensitive to the generation of a transmembrane delta pH. We carried out a kinetic analysis of each enzyme complex studying the proton translocation and the electron transfer activities of complex I as well as the proton translocation and the ATP hydrolytic activities of complex V. A micromethod to prepare coupled submitochondrial particles from platelets might be useful to investigate cell bioenergetic damage occurring in mitochondrial diseases and ageing.
Baracca
A
A
Department of Biochemistry G. Moruzzi, Bologna, Italy.
Bucchi
L
L
Ghelli
A
A
Lenaz
G
G
D000251:Adenosine Triphosphatases / Q000097:blood*; D000818:Animals; D001792:Blood Platelets / Q000201:enzymology*; D002352:Carrier Proteins; D004579:Electron Transport; D006736:Horses; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000502:physiology; D007700:Kinetics; D008565:Membrane Proteins / Q000097:blood*; D008832:Microchemistry; D025261:Mitochondrial Proton-Translocating ATPases; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000097:blood*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013367:Submitochondrial Particles / Q000201:enzymology*
D002352:Carrier Proteins; D008565:Membrane Proteins; D016660:NAD(P)H Dehydrogenase (Quinone); D000251:Adenosine Triphosphatases; D025261:Mitochondrial Proton-Translocating ATPases; C020439:oligomycin sensitivity-conferring protein