Molecular processes of inhibition and stimulation of ATP synthase caused by the phytotoxin tentoxin.
F1-ATPase is the smallest mechanical motor known. Tentoxin, a cyclic peptide produced by phytopathogenic fungi, inactivates the F1 motor in sensitive plants at nanomolar to micromolar concentrations, whereas higher concentrations surpass the natural activity of the enzyme. Single molecule studies now have clarified the molecular steps involved in both processes. Inactivation delays the dwell time of a single step in the complete 360 degrees turn and results in an asymmetric rotation of the central rotor subunit. In contrast, rotation in the stimulated F1 particle is smooth and accompanied by strongly reduced ADP inhibition. Our study provides for the first time the direct observation of a noncompetitively inhibited state of the enzyme and directly visualizes the regulation of the molecular motor by an external natural compound. In addition, the ADP release step during catalysis was revealed by analysis of the single molecule rotation behavior. Hence, tentoxin is a sophisticated molecular tool to mark and control certain catalytic steps within the reaction pathway of the molecular F1 motor.
Meiss
Erik
E
Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259-R1-8, Yokohama 226-8503, Japan.
Konno
Hiroki
H
Groth
Georg
G
Hisabori
Toru
T
D000244:Adenosine Diphosphate / Q000737:chemistry / Q000378:metabolism; D002384:Catalysis; D004789:Enzyme Activation; D020409:Molecular Motor Proteins / Q000737:chemistry* / Q000378:metabolism; D009183:Mycotoxins / Q000737:chemistry* / Q000378:metabolism; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000378:metabolism; D010944:Plants / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000378:metabolism
D020409:Molecular Motor Proteins; D009183:Mycotoxins; D010456:Peptides, Cyclic; D010940:Plant Proteins; D000244:Adenosine Diphosphate; D006180:Proton-Translocating ATPases; C003384:tentoxin