The regulator of the F1 motor: inhibition of rotation of cyanobacterial F1-ATPase by the epsilon subunit.
The chloroplast-type F(1) ATPase is the key enzyme of energy conversion in chloroplasts, and is regulated by the endogenous inhibitor epsilon, tightly bound ADP, the membrane potential and the redox state of the gamma subunit. In order to understand the molecular mechanism of epsilon inhibition, we constructed an expression system for the alpha(3)beta(3)gamma subcomplex in thermophilic cyanobacteria allowing thorough investigation of epsilon inhibition. epsilon Inhibition was found to be ATP-independent, and different to that observed for bacterial F(1)-ATPase. The role of the additional region on the gamma subunit of chloroplast-type F(1)-ATPase in epsilon inhibition was also determined. By single molecule rotation analysis, we succeeded in assigning the pausing angular position of gamma in epsilon inhibition, which was found to be identical to that observed for ATP hydrolysis, product release and ADP inhibition, but distinctly different from the waiting position for ATP binding. These results suggest that the epsilon subunit of chloroplast-type ATP synthase plays an important regulator for the rotary motor enzyme, thus preventing wasteful ATP hydrolysis.
Konno
Hiroki
H
Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta, Yokohama, Japan.
Murakami-Fuse
Tomoe
T
Fujii
Fumihiko
F
Koyama
Fumie
F
Ueoka-Nakanishi
Hanayo
H
Pack
Chan-Gi
CG
Kinjo
Masataka
M
Hisabori
Toru
T
D000255:Adenosine Triphosphate / Q000378:metabolism; D000458:Cyanobacteria / Q000201:enzymology*; D004123:Dimethylamines / Q000494:pharmacology; D006868:Hydrolysis / Q000187:drug effects; D020409:Molecular Motor Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D050505:Mutant Proteins / Q000378:metabolism; D011485:Protein Binding; D021122:Protein Subunits / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D012399:Rotation
D004123:Dimethylamines; D020409:Molecular Motor Proteins; D050505:Mutant Proteins; D021122:Protein Subunits; C014518:dodecyldimethylamine oxide; D000255:Adenosine Triphosphate; C444660:gamma subunit, F(1) ATPase; D006180:Proton-Translocating ATPases