Deleting the IF-like subunit from ATP synthase is not sufficient to activate ATP hydrolysis.

In oxidative phosphorylation, ATP synthases interconvert two forms of free energy: they are driven by the proton-motive force across an energy-transducing membrane to synthesize ATP and displace the ADP/ATP ratio from equilibrium. For thermodynamically efficient energy conversion they must be reversible catalysts. However, in many species ATP synthases are unidirectional catalysts (their rates of ATP hydrolysis are negligible), and in others mechanisms have evolved to regulate or minimize hydrolysis. Unidirectional catalysis by ATP synthase has been attributed to its unique subunit, which is structurally analogous to the mammalian inhibitor protein IF Here, we used homologous recombination to delete the subunit from the genome, and compared ATP synthesis and hydrolysis by the wild-type and knockout enzymes in inverted membrane vesicles and the F-ATPase subcomplex. ATP synthesis was not affected by loss of the subunit, and the rate of ATP hydrolysis increased by less than twofold, remaining negligible in comparison with the rates of the and mammalian enzymes. Therefore, deleting the subunit is not sufficient to activate ATP hydrolysis. We close by considering our conclusions in the light of reversible catalysis and regulation in ATP synthase enzymes.