F-1-ATPase, the C-terminal end of subunit gamma is not required for ATP hydrolysis-driven rotation
Autor(en): | Muller, M Panke, O Junge, W Engelbrecht, S |
Stichwörter: | BETA-SUBUNIT; Biochemistry & Molecular Biology; CATALYSIS; ESCHERICHIA-COLI F1-ATPASE; NONCATALYTIC SITES; ROTARY F-ATPASE; SITE-DIRECTED MUTAGENESIS; SYNTHASE; TORQUE; TRANSPORT; VISCOELASTIC DYNAMICS | Erscheinungsdatum: | 2002 | Herausgeber: | AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC | Journal: | JOURNAL OF BIOLOGICAL CHEMISTRY | Volumen: | 277 | Ausgabe: | 26 | Startseite: | 23308 | Seitenende: | 23313 | Zusammenfassung: | ATP hydrolysis by the isolated F-1-ATPase drives the rotation of the central shaft, subunit gamma, which is located within a hexagon formed by subunits (alphabeta)(3). The C-terminal end of gamma forms an alpha-helix which properly fits into the ``hydrophobic bearing'' provided by loops of subunits alpha and beta. This ``bearing'' is expected to be essential for the rotary function. We checked the importance of this contact region by successive C-terminal deletions of 3, 6, 9, 12, 15, and 18 amino acid residues (Escherichia coli F-1-ATPase). The ATP hydrolysis activity of a load-free ensemble of F-1 with 12 residues deleted decreased to 24%, of the control. EF1 with deletions of 15 or 18 residues was inactive, probably because it failed to assemble. The average torque generated by a single molecule of EF1 when loaded by a fluorescent actin filament was, however, unaffected by deletions of up to 12 residues, as was their rotational behavior (all samples rotated during 60 /- 19% of the observation time). Activation energy analysis with the ensemble revealed a moderate decrease from 54 kJ/mol for EF1 (full-length gamma) to 34 kJ/mol for EF1(gamma-12). These observations imply that the intactness of the C terminus of subunit gamma provides structural stability and/or routing during assembly of the enzyme, but that it is not required for the rotary action under load, proper. |
ISSN: | 00219258 | DOI: | 10.1074/jbc.M201998200 |
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geprüft am 06.05.2024