The effect of mild trypsin digestion of F1 on energy coupling in the mitochondrial ATP synthase.

Mild trypsin digestion of isolated bovine-heart mitochondrial F1-ATPase removed the first 15 residues from the N-terminus of subunit alpha under conditions in which other F1 subunits were apparently untouched. When the trypsinized F1 (TF1) was reconstituted with the F0 sector in the mitochondrial membrane (USMP), the ATP hydrolase activity acquired oligomycin sensitivity but ATP hydrolysis was decoupled from proton pumping. TF1 added to USMP did not block the proton channel in F0 as the native F1 did. AMP-PNP inhibited proton conductivity in reconstituted F1-USMP but this effect was lost in reconstituted TF1-USMP. These results indicate that the N-terminus of the F1 alpha subunit plays a critical role in the conformational communication between F1 and F0.