ATP synthase F(1) sector rotation. Defective torque generation in the beta subunit Ser-174 to Phe mutant and its suppression by second mutations.
Subunit gamma of the ATP synthase F(1) sector is located at the center of the alpha(3)beta(3) hexamer and rotates unidirectionally during ATP hydrolysis, generating the rotational torque of approximately 45 pN.nm. A mutant F(1) with the betaSer-174 to Phe substitution (betaS174F) in the beta subunit generated lower torque ( approximately 17 pN.nm), indicating that betaS174F is mechanically defective, the first such mutant reported. The defective rotation of betaS174F was suppressed by a second-site mutation, betaGly-149 to Ala, betaIle-163 to Ala, or betaIle-166 to Ala in the same subunit, but not by betaLeu-238 to Ala. These results suggest that the region between betaGly-149 and betaSer-174 plays an important role in the coupling between ATP hydrolysis and mechanical work.
Iko
Y
Y
Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, CREST (Core Research for Evolutional Science and Technology) of Japan Science and Technology Corporation, Ibaraki, Osaka 567-0047, Japan.
Sambongi
Y
Y
Tanabe
M
M
Iwamoto-Kihara
A
A
Saito
K
K
Ueda
I
I
Wada
Y
Y
Futai
M
M
D000199:Actins / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000378:metabolism; D000409:Alanine / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology; D005998:Glycine / Q000737:chemistry; D006868:Hydrolysis; D007532:Isoleucine / Q000737:chemistry; D008958:Models, Molecular; D009154:Mutation; D010649:Phenylalanine / Q000737:chemistry; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D012694:Serine / Q000737:chemistry*; D013997:Time Factors
D000199:Actins; D007532:Isoleucine; D012694:Serine; D010649:Phenylalanine; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases; D000409:Alanine; D005998:Glycine