Quaternary structure of chloroplast F1-ATPase in solution. Conformational changes in spatial arrangement of subunits upon activation.
The hydrodynamic properties of isolated ATPases were studied via their rotational diffusion in buffer solution. Chloroplast F1-ATPase (CF1) and Escherichia coli F1-ATPase (EF1) were covalently labeled with eosinisothiocyanate and then investigated by polarized laser spectroscopy. The rotational correlation time in aqueous buffer of latent (five-subunit) CF1 was 390 ns. Four-subunit (delta-deficient) CF1 showed the same correlation time, however, for three-subunit (delta, epsilon-deficient) CF1 the rotational correlation time was more than eight times larger (3200 ns). The rotational correlation time of activated CF1 was three times larger than the one of latent CF1. These large changes in the rotational correlation times are directly related to changes in the quaternary structure of CF1 upon activation. EF1 was found to behave essentially as activated CF1. Based on the observed rotational correlation times we concluded that the mass distributions of latent CF1 and of delta-deficient CF1 resemble a dimeric arrangement. The structure of delta, epsilon-deficient CF1 more likely resembles a hexagon, the mass centers of the six main subunits lie in one plane. The structure of the activated forms of CF1 can be described best as an intermediate between the dimeric arrangement of latent CF1 and an octahedron. The large changes in the quaternary structure of isolated CF1 are reversed when the activation of the enzyme is reversed.
Wagner
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Engelbrecht
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Andreo
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D055598:Chemical Phenomena; D002621:Chemistry; D002627:Chemistry, Physical; D002736:Chloroplasts / Q000201:enzymology*; D004058:Diffusion; D004789:Enzyme Activation; D007834:Lasers; D046911:Macromolecular Substances; D008433:Mathematics; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000378:metabolism; D012399:Rotation; D012996:Solutions; D013057:Spectrum Analysis
D046911:Macromolecular Substances; D012996:Solutions; D006180:Proton-Translocating ATPases