|Title||Structure of the γ-ε complex of cyanobacterial F-ATPase reveals a suppression mechanism of the γ subunit on ATP hydrolysis in phototrophs.|
|Publication Type||Journal Article|
|Year of Publication||2018|
|Authors||Murakami, S, Kondo, K, Katayama, S, Hara, S, Sunamura, E-I, Yamashita, E, Groth, G, Hisabori, T|
|Date Published||2018 09 18|
|Keywords||Adenosine Triphosphate, Bacterial Proteins, Crystallography, X-Ray, Cyanobacteria, Hydrolysis, Protein Structure, Secondary, Proton-Translocating ATPases|
F-ATPase forms the membrane-associated segment of FF-ATP synthase - the fundamental enzyme complex in cellular bioenergetics for ATP hydrolysis and synthesis. Here, we report a crystal structure of the central F subcomplex, consisting of the rotary shaft γ subunit and the inhibitory ε subunit, from the photosynthetic cyanobacterium 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 subcomplex. Biochemical analyses of the corresponding αβγ complex confirm that the clinch of the hairpin structure plays a critical role and accounts for a significant interaction in the αβ 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 rotation during ATP hydrolysis.
|Alternate Journal||Biochem. J.|