Reversible ischemic inhibition of F(1)F(0)-ATPase in rat and human myocardium.
The physiological role of F(1)F(0)-ATPase inhibition in ischemia may be to retard ATP depletion although views of the significance of IF(1) are at variance. We corroborate here a method for measuring the ex vivo activity of F(1)F(0)-ATPase in perfused rat heart and show that observation of ischemic F(1)F(0)-ATPase inhibition in rat heart is critically dependent on the sample preparation and assay conditions, and that the methods can be applied to assay the ischemic and reperfused human heart during coronary by-pass surgery. A 5-min period of ischemia inhibited F(1)F(0)-ATPase by 20% in both rat and human myocardium. After a 15-min reperfusion a subsequent 5-min period of ischemia doubled the inhibition in the rat heart but this potentiation was lost after 120 min of reperfusion. Experiments with isolated rat heart mitochondria showed that ATP hydrolysis is required for effective inhibition by uncoupling. The concentration of oligomycin for 50% inhibition (I(50)) for oxygen consumption was five times higher than its I(50) for F(1)F(0)-ATPase. Because of the different control strengths of F(1)F(0)-ATPase in oxidative phosphorylation and ATP hydrolysis an inhibition of the F(1)F(0)-ATPase activity in ischemia with the resultant ATP-sparing has an advantage even in an ischemia/reperfusion situation.
Department of Medical Biochemistry, University, of Oulu, Finland.
K
K
Ylitalo
A
A
Ala-Rämi
K
K
Vuorinen
K
K
Peuhkurinen
M
M
Lepojärvi
P
P
Kaukoranta
K
K
Kiviluoma
I
I
Hassinen
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001706:Biopsy; D006348:Cardiac Surgical Procedures; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008297:Male; D008929:Mitochondria, Heart / Q000187:drug effects / Q000201:enzymology; D017202:Myocardial Ischemia / Q000201:enzymology* / Q000601:surgery; D009206:Myocardium / Q000201:enzymology; D009840:Oligomycins / Q000494:pharmacology; D010085:Oxidative Phosphorylation; D010477:Perfusion; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors*; D051381:Rats; D017207:Rats, Sprague-Dawley
D009840:Oligomycins; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases