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• W2044480820 abstract "Conditions for the dissociation and reassembly of the multi-subunit vacuolar proton-translocating ATPase (H+-ATPase) from oat roots (Avena sativa var Lang) were investigated. The peripheral sector of the vacuolar H+-ATPase is dissociated from the membrane integral sector by chaotropic anions. Membranes treated with 0.5 molar KI lost 90% of membrane-bound ATP hydrolytic activity; however, in the presence of Mg2+ and ATP, only 0.1 molar KI was required for complete inactivation of ATPase and H+-pumping activities. A high-affinity binding site for MgATP (dissociation constant = 34 micromolar) was involved in this destabilization. The relative loss of ATPase activity induced by KI, KNO3, or KCl was accompanied by a corresponding increase in the peripheral subunits in the supernatant, including the nucleotide-binding polypeptides of 70 and 60 kilodaltons. The order of effectiveness of the various ions in reducing ATPase activity was: KSCN > KI > KNO3 > KBr > K-acetate > K2SO4 > KCl. The specificity of nucleotides (ATP > GTP > ITP) in dissociating the ATPase is consistent with the participation of a catalytic site in destabilizing the enzyme complex. Following KI-induced dissociation of the H+-ATPase, the removal of KI and MgATP by dialysis resulted in restoration of activity. During dialysis for 24 hours, ATP hydrolysis activity increased to about 50% of the control. Hydrolysis of ATP was coupled to H+ pumping as seen from the recovery of H+ transport following 6 hours of dialysis. Loss of the 70 and 60 kilodalton subunits from the supernatant as probed by monoclonal antibodies further confirmed that the H+-ATPase complex had reassembled during dialysis. These data demonstrate that removal of KI and MgATP resulted in reassociation of the peripheral sector with the membrane integral sector of the vacuolar H+-ATPase to form a functional H+ pump. The ability to dissociate and reassociate in vitro may have implications for the regulation, biosynthesis, and assembly of the vacuolar H+-ATPase in vivo." @default.
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• W2044480820 date "1992-05-01" @default.
• W2044480820 modified "2023-09-27" @default.
• W2044480820 title "Dissociation and Reassembly of the Vacuolar H+-ATPase Complex from Oat Roots" @default.
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• W2044480820 doi "https://doi.org/10.1104/pp.99.1.161" @default.
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