title: Energy-dependent changes in conformation and catalytic activity of the chloroplast ATP synthase.
abstract: Energy-dependent activation of the chloroplast ATP synthase (CF0CF1) has been elucidated by investigating the conformational changes, the ADP effect, and the catalytic cooperativity of ATP hydrolysis. Conformational change was observed by measuring the reactivity of Lys-109 of the epsilon subunit of chloroplast coupling factor 1 with pyridoxal 5'-phosphate. In the postillumination dark, the Lys-109 reactivity decreased biphasically with half-times of less than 1 and 17 s. NH4Cl accelerated the slow phase decrease. Addition of ADP (0.2 microM) in the postillumination dark inactivated CF0CF1 (0.05 microM) with a half-time of 12 s. At high concentration of CF0CF1 (1.2 microM), inactivation occurred without exogenously added ADP with a half-time of 12 s. Accompanying the inactivation, the positive catalytic cooperativity of ATP hydrolysis decreased. Addition of 10 mM NH4Cl before ADP (0.2 microM) decelerated the ADP-induced inactivation to a half-time of 64 s. Throughout this inactivation, the positive catalytic cooperativity was maintained at a high level. These results suggest three distinct conformations of CF0CF1, EH, EM, and EL, and their ADP binding forms EM-ADP and EL-ADP. EH, EM, and EL have a low affinity for ADP, a high affinity for ADP, and low accessibility to ADP, respectively. EM and EL exhibit highly cooperative ATP hydrolysis. ATP hydrolysis catalyzed by EM-ADP exhibits no cooperativity. EL-ADP is inactive.
authors.affiliation: Department of Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Kanagawa, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Komatsu-Takaki
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors; D000595:Amino Acid Sequence; D002384:Catalysis; D002736:Chloroplasts / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D004734:Energy Metabolism; D006868:Hydrolysis; D008969:Molecular Sequence Data; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000378:metabolism*; D011732:Pyridoxal Phosphate / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
chemical_list: D011732:Pyridoxal Phosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D014357:Trypsin; D006180:Proton-Translocating ATPases