Structure of the γ-ε complex of cyanobacterial F 1 -ATPase reveals a suppression mechanism of the γ subunit on ATP hydrolysis in phototrophs.

F ₁ -ATPase forms the membrane-associated segment of F ₀ F ₁ -ATP synthase - the fundamental enzyme complex in cellular bioenergetics for ATP hydrolysis and synthesis. Here, we report a crystal structure of the central F ₁ subcomplex, consisting of the rotary shaft γ subunit and the inhibitory ε subunit, from the photosynthetic cyanobacterium Thermosynechococcus elongatus BP-1, at 1.98 Å resolution. In contrast with their homologous bacterial and mitochondrial counterparts, the γ subunits of photosynthetic organisms harbour a unique insertion of 35-40 amino acids. Our structural data reveal that this region forms a β-hairpin structure along the central stalk. We identified numerous critical hydrogen bonds and electrostatic interactions between residues in the hairpin and the rest of the γ subunit. To elaborate the critical function of this β-hairpin in inhibiting ATP hydrolysis, the corresponding domain was deleted in the cyanobacterial F ₁ subcomplex. Biochemical analyses of the corresponding α ₃ β ₃ γ complex confirm that the clinch of the hairpin structure plays a critical role and accounts for a significant interaction in the α ₃ β ₃ complex to induce ADP inhibition during ATP hydrolysis. In addition, we found that truncating the β-hairpin insertion structure resulted in a marked impairment of the interaction with the ε subunit, which binds to the opposite side of the γ subunit from the β-hairpin structure. Combined with structural analyses, our work provides experimental evidence supporting the molecular principle of how the insertion region of the γ subunit suppresses F ₁ rotation during ATP hydrolysis.
(© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)