ATP hydrolysis-driven H(+) translocation is stimulated by sulfate, a strong inhibitor of mitochondrial ATP synthesis.
Sulfate is a partial inhibitor at low and a non-essential activator at high [ATP] of the ATPase activity of F(1). Therefore, a catalytically-competent ternary F(1) x ATP x sulfate complex can be formed. In addition, the ANS fluorescence enhancement driven by ATP hydrolysis in submitochondrial particles is also stimulated by sulfate, clearly showing that the ATP hydrolysis in its presence is coupled to H(+) translocation. However, sulfate is a strong linear inhibitor of the mitochondrial ATP synthesis. The inhibition was competitive (K (i) = 0.46 mM) with respect to Pi and mixed (K (i) = 0.60 and K'(i) = 5.6 mM) towards ADP. Since it is likely that sulfate exerts its effects by binding at the Pi binding subdomain of the catalytic site, we suggest that the catalytic site involved in the H(+) translocation driven by ATP hydrolysis has a more open conformation than the half-closed one (beta(HC)), which is an intermediate in ATP synthesis. Accordingly, ATP hydrolysis is not necessarily the exact reversal of ATP synthesis.
Sección Biología Molecular, Instituto de Biología Molecular y Celular de Rosario (IBR), CONICET-Universidad Nacional de Rosario, Rosario, Argentina. lodeyro@ibr.gov.ar
Anabella F
AF
Lodeyro
María V
MV
Castelli
Oscar A
OA
Roveri
D000255:Adenosine Triphosphate / Q000138:chemical synthesis* / Q000737:chemistry*; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D003198:Computer Simulation; D004789:Enzyme Activation; D004795:Enzyme Stability; D006868:Hydrolysis; D008929:Mitochondria, Heart / Q000201:enzymology*; D008956:Models, Chemical; D008958:Models, Molecular; D009206:Myocardium / Q000201:enzymology*; D011485:Protein Binding; D011487:Protein Conformation; D021122:Protein Subunits / Q000737:chemistry; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000648:ultrastructure*; D011522:Protons; D013431:Sulfates / Q000737:chemistry*
D021122:Protein Subunits; D011522:Protons; D013431:Sulfates; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases