biostudies-literatureMao HZNIGMS NIH HHS18478-83https://www.ebi.ac.uk/biostudies/studies/S-EPMC2141802biostudies-literatureUnknown104(47)ATP synthase uses a unique rotary mechanism to couple ATP synthesis and hydrolysis to transmembrane proton translocation. The F(1) subcomplex has three catalytic nucleotide binding sites, one on each beta subunit, with widely differing affinities for MgATP or MgADP. During rotational catalysis, the sites switch their affinities. The affinity of each site is determined by the position of the central gamma subunit. The site with the highest nucleotide binding affinity is catalytically active. From the available x-ray structures, it is not possible to discern the high-affinity site. Using fluorescence resonance energy transfer between tryptophan residues engineered into gamma and trinitrophenyl nucleotide analogs on the catalytic sites, we were able to determine that the high-affinity site isProceedings of the National Academy of Sciences of the United States of AmericaIdentification of the betaTP site in the x-ray structure of F1-ATPase as the high-affinity catalytic site.PMC2141802GM071462R01 GM071462Weber JMao HZfalseIdentification of the betaTP site in the x-ray structure of F1-ATPase as the high-affinity catalytic site.ATP synthase uses a unique rotary mechanism to couple ATP synthesis and hydrolysis to transmembrane proton translocation. The F(1) subcomplex has three catalytic nucleotide binding sites, one on each beta subunit, with widely differing affinities for MgATP or MgADP. During rotational catalysis, the sites switch their affinities. The affinity of each site is determined by the position of the central gamma subunit. The site with the highest nucleotide binding affinity is catalytically active. From the available x-ray structures, it is not possible to discern the high-affinity site. Using fluorescence resonance energy transfer between tryptophan residues engineered into gamma and trinitrophenyl nucleotide analogs on the catalytic sites, we were able to determine that the high-affinity site is2007-01-01T00:00:00Z2007 Nov2025-04-18T19:32:05.697Z2019-03-27T02:22:09ZS-EPMC21418021800389610.1073/pnas.0709322104