Functional Reconstitution of Plasma Membrane H^+-ATPase from Mung Bean ( Vigna radiata L.) Hypocotyls in Liposomes Prepared with Various Molecular Species of Phospholipids

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概要

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• The activity of solubilized plasma membrane ATPase is affected by the nature of exogenously added molecular species of phospholipids. To examine the role of the polar head group and of the molecular species of phospholipids in H^+-pumping, the ATPase solubilized from plasma membranes of mung bean (Vigna radiata L.) hypocotyls was reconstituted in liposomes prepared with a variety of phospholipids. The extent of activation of solubilized plasma membrane ATPase due to the addition of 1-palmitoyl 2-oleoyl-phospholipids (PO-phospholipids) and asolectin decreased in the following order: POPS > POPC ≩ asolectin ≩ POPG > POPE > POPA (see List of Abbreviations). H^+-pumping into proteoliposomes reconstituted with asolectin and plasma membrane ATPase was demonstrated by quinacrine fluorescence quenching in the presence of ATP-MgSO_4. H^+-pumping was inhibited by VO_4 and gramicidin D. When plasma membrane ATPase was reconstituted in liposomes prepared with various PO-phospholipids, the ability of PO-phospho-lipids to support H^+-pumping into the proteoliposomes decreased in the following order: POPG ≩ POPS > asolectin ≩ POPC. POPE and POPA failed to support any H^+-pumping. A remarkably high rate of H^+-pumping was observed in proteoliposomes prepared with 1-saturated 2-unsaturated fatty acids, such as POPC, but H^+-pumping could hardly be detected in proteoliposomes prepared with 1-, 2-unsaturated or 1-, 2-saturated fatty acids, such as PSPC or DLPC. ATPase activity in proteoliposomes was dependent on the species of PO-phospholipids used for reconstitution and decreased in the following order: POPS > POPG > POPC ≩ asolectin > POPA > POPE. DLPC (see List of Abbreviations) which includes a 1-, 2-unsaturated fatty acid supported only markedly depressed activity. Both H^+-pumping and the hydrolysis of ATP by the plasma membrane ATPase are strongly affected by the polar head group and composition of the fatty acyl chain of phospholipids used to prepare liposomes for reconstitution of the ATPase.

• 日本植物生理学会の論文

著者

• Kasamo Kunihiro

  Molecular Function Laboratory Biological Function Division National Food Research Institute

• Kasamo Kunihiro

  Molecular Function Laboratory National Food Research Institute

• Yamanishi Hiroyasu

  Molecular Function Laboratory Biological Function Division National Food Research Institute

• Yamanishi Hiroyasu

  Molecular Function Laboratory National Food Research Institute

関連論文

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• Modulation of the Activity of Purified Tonoplast H^+-ATPase from Mung Bean (Vigna radiata L.) Hypocotyls by Various Lipids
• Low Temperature-Induced Changes in the Thermotropic Properties and Fatty Acid Composition of the Plasma Membrane and Tonoplast of Cultured Rice (Oryza sativa L.) Cells : ENVIRONMENTAL AND STRESS RESPONSES : MEMBRANES AND BIOENERGETICS
• Functional Reconstitution of Plasma Membrane H^+-ATPase from Mung Bean ( Vigna radiata L.) Hypocotyls in Liposomes Prepared with Various Molecular Species of Phospholipids
• Effects of Cerebroside and Cholesterol on the Reconstitution of Tonoplast H^+-ATPase Punfied from Mung Bean(Vigna radiata L.)Hypocotyls in Liposomes : MEMBRANES AND BIOENERGETICS

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