Reconstitution and Characterization of Plasma Membrane H^+-ATPase Activity from Rice Roots

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概要

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• Plasma membrane vesicles were purified from rice (Oryza sativa L. cv. Nipponbare) roots using aqueous two-phase partitioning. The purified vesicles were solubilized with deoxycholate and reconstituted with soybean phospholipids by gel filtration. Ionophores simulated the ATPase activity of the reconstituted vesicles. Inhibition of the ATPase activity by vanadate increased from 77 to 93% by the reconstitution. These results indicate that the reconstituted vesicles were sealed and the vanadate-sensitive ATPase activity was purified during the reconstitution. The ATPase activity of the reconstituted vesicles was characterized. Only ATP was effectively hydrolyzed. Apparent K_m value was 1.26 mM. Divalent cations were necessary for the activity, and their stimulation was in the following order : Mg^<2+> > Fe^<2+> > Mn^<2+>, Co^<2+> ≫ Ca^<2+>. Half-maximal inhibition by vanadate occurred at μM. Monovalent ions stimulated the activity, and cations and anions were effective in the order of NH_4^+ > K^+, Rb^+ > Na^+ > Li^+ > choline^+ and NO_3^- > Cl^-, Br^- > I^- > SO_4^<2->, respectively. Optimum pH was 6.5.

• 社団法人日本土壌肥料学会の論文

著者

• SUEYOSHI Kuni

  Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University

• WAKIUCHI Nariaki

  Department of Agricultural Chemistry, Faculty of Agriculture, Kobe University

• HARADA Hisatomi

  Department of Agricultural Chemistry, Faculty of Agriculture, Kobe University

• OJI Yoshikiyo

  Department of Agricultural Chemistry, Faculty of Agriculture, Kobe University

• Sueyoshi Kuni

  Department Of Agricultural Chemistry Faculty Of Agriculture Kobe University

• Oji Yoshikiyo

  Department Of Agricultural Chemistry Faculty Of Agriculture Kobe University

• Harada H

  National Institute Of Livestock And Grassland Science:(present Office)agriculture Forestry And Fishe

• Wakiuchi Nariaki

  Department Of Agricultural Chemistry Faculty Of Agriculture Kobe University

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