Movie S3.

Generation of a 30° rotation of the γ-subunit in bovine F1-ATPase by release of phosphate from the βE-subunit. The movie commences with a view from the side of F1-ThioP in ribbon representation, with the bound thiophosphate shown as orange spheres. The α- and β-subunits are shown in red and yellow, respectively. Then, the αTP-, αDP-, βTP-, and βDP-subunits are removed sequentially, leaving the αEβE-dimer and the central stalk subunits γ, δ, and ε (blue, magenta, and green, respectively). This residual subcomplex is turned to the right around the y axis by 100° to expose a side view of the αE-subunit and the central stalk. Then, the δ- and ε-subunits and residues 42–210 of the γ-subunit are removed, and the βE-subunit becomes fainter. In the final frames, the bound phosphate is released and the αE-subunit opens and moves away from the γ-subunit, disrupting packing interactions between the αE- and γ-subunits. To reestablish these interactions, the γ-subunit rotates by 30°. The structure of the final state is the postphosphate release state of bovine F1-ATPase found in F1-I3-ThioP.