1998 Fiscal Year Annual Research Report

植物細胞質膜と液胞膜の水チャネルの分子特性と細胞機能の解析

Research Project

Project/Area Number	09660114
Research Institution	Nagoya University
Principal Investigator	前島正義名古屋大学, 農学部, 助教授 (80181577)
Keywords	細胞膜 / 液胞膜 / 水チャネル / 分子多様性 / ダイコン / GFP
Research Abstract	水チャネルは1990年代になって発見された膜輸送分子で、浸透圧に応答した水の膜透過を促進するチャネル分子である。動物、植物に共通して存在する。本研究ではダイコンを主研究材料として、細胞膜と液胞膜の水チャネルについて、多数の分子種の一次構造の特徴および細胞機能を解明することを研究課題とした。具体的には次の成果を得た。(1)ダイコンから液胞膜型(γ-,δ-VM23)、細胞膜型(PAQ1,1b,1c,2,2b,2c)の水チャネルcDNAを得た。細胞膜型は液胞膜型よりもN端側が20-30残基長く、一次構造上の相同性は低い(identity,40%)。(2)いずれの水チャネルもダイコン実生の胚軸、生長中の根・葉などでmRNAが多く蓄積しており、細胞の生長時に活発に合成されていた。これ以外の組織においては、各分子種は独自の発現パターンを示し、個別に発現が調節されているものと推定された。(3)特に液胞膜水チャネルは量が多く、最も多い場合液胞膜タンパク質の40%を占める。その60%以上はγ-VM23であること、さらにダイコンの細胞膜では約10%に達することが明らかになった。本研究では、水チャネルの細胞膜、液胞膜への局在化機構を解明することを目的に、green fluorescent protein(GFP)と水チャネル分子の融合タンパク質をタバコ培養細胞BY-2で発現させることを試みた。いずれの水チャネル分子も目的の膜に局在することを確認できた。本研究により、水チャネル分子を通して植物の成長、組織分化、水生理現象を、今後さらに解明していくための基礎的知見と実験系が整った。

Research Products
(15 results)

All Other

All Publications (15 results)

[Publications] Fleurat-Lessard,P.: "Increased expression of vacuolar aquaporin and H^+-AT Pase related to motor cell function in Mimosa pudica L." Plant Physiology. 114. 827-834 (1997)
[Publications] Shiratake,K.: "Changes in H^+-pumps and a tonoplast intrinsic protein of vacuolar membranes during the development of pear fruit." Plant & Cell Physiology. 38. 1039-1045 (1997)
[Publications] Takasu,A.: "Analysis of substrate binding site and carboxyl terminal region of vacuolar H^+-translocating pyrophoshatase of mung bean with peptide antibodies." Journal of Biochemistry. 122. 883-889 (1997)
[Publications] Nakanishi,Y.: "Molecular cloning of vacuolar H^+-pyrophosphtase and its developmental expression in growing hypocotyl of mung bean." Plant Physiology. 116. 589-597 (1998)
[Publications] Smart,L.B.: "Genes involved in osmoregulation during turgor-driven cell expansion of developing cotton fibers are differentially regulated." Plant Physiology. 116. 1539-1549 (1998)
[Publications] Kasai,M.: "The activity of the root vacuolar H^+-pyrophosphatase in rye plants grown under deficient conditions of mineral nutrients." Plant & Cell Physiology. 39. 890-894 (1998)
[Publications] Shiratake,K.: "Changes in tonoplast protein and density with the development of pear fruit." Physiologia Plantrum. 103. 312-319 (1998)
[Publications] Higuchi,T.: "Molecular cloning, water channel activity and tissue specific expression of two isoforms of radish vacuolar aquaporin." Plant & Cell Physiology. 39. 905-913 (1998)
[Publications] Serraj,R.: "Highly expressed vacuolar aquaporin in the crotex of Glycine max nitrogen-fixing nodules." Planta. 206. 681-684 (1998)
[Publications] Ikeda,M.: "A vacuolar inorganic H^+-pyrophosphatase in Acetabularia acetabulum : Partial purification, characterization and molecular cloning." Journal of Experimental Biology. 50. 139-140 (1998)
[Publications] Sasaki,T.: "Development of vacuoles and vacuolar ATPase activity under extremely high-CO_2 conditions in Chlorococcum littorale cells." Plant Biology. 1(印刷中). (1999)
[Publications] Ueoka-Nakanishi,H.: "Properties and molecular cloning of Ca^<2+>/H^+ antiporter in the vacuolar membrane of mung bean." European Journal of Biochemistry. 印刷中. (1999)
[Publications] Maeshima,M.: "Cycloprodigiosin uncouples H^+-pyrophosphatase of plant vacuolar membrane in the presence of chloride ion." Plant & Cell Physiology. 印刷中. (1999)
[Publications] Nakanishi,Y.: "Molecular cloning and sequencing of the cDNA for vacuolar H^+-pyrophosphatase from Chara corallina." Biochimica et Biophysica Acta. 印刷中. (1999)
[Publications] 前島正義: "細胞工学別冊「植物のタンパク質実験」" 液胞膜H^+-ピロホスファターゼの精製, 131-135 (1998)

1998 Fiscal Year Annual Research Report

植物細胞質膜と液胞膜の水チャネルの分子特性と細胞機能の解析

Principal Investigator

前島 正義 名古屋大学, 農学部, 助教授 (80181577)

Research Products

[Publications] Fleurat-Lessard,P.: "Increased expression of vacuolar aquaporin and H^+-AT Pase related to motor cell function in Mimosa pudica L." Plant Physiology. 114. 827-834 (1997)

[Publications] Shiratake,K.: "Changes in H^+-pumps and a tonoplast intrinsic protein of vacuolar membranes during the development of pear fruit." Plant & Cell Physiology. 38. 1039-1045 (1997)

[Publications] Takasu,A.: "Analysis of substrate binding site and carboxyl terminal region of vacuolar H^+-translocating pyrophoshatase of mung bean with peptide antibodies." Journal of Biochemistry. 122. 883-889 (1997)

[Publications] Nakanishi,Y.: "Molecular cloning of vacuolar H^+-pyrophosphtase and its developmental expression in growing hypocotyl of mung bean." Plant Physiology. 116. 589-597 (1998)

[Publications] Smart,L.B.: "Genes involved in osmoregulation during turgor-driven cell expansion of developing cotton fibers are differentially regulated." Plant Physiology. 116. 1539-1549 (1998)

[Publications] Kasai,M.: "The activity of the root vacuolar H^+-pyrophosphatase in rye plants grown under deficient conditions of mineral nutrients." Plant & Cell Physiology. 39. 890-894 (1998)

[Publications] Shiratake,K.: "Changes in tonoplast protein and density with the development of pear fruit." Physiologia Plantrum. 103. 312-319 (1998)

[Publications] Higuchi,T.: "Molecular cloning, water channel activity and tissue specific expression of two isoforms of radish vacuolar aquaporin." Plant & Cell Physiology. 39. 905-913 (1998)

[Publications] Serraj,R.: "Highly expressed vacuolar aquaporin in the crotex of Glycine max nitrogen-fixing nodules." Planta. 206. 681-684 (1998)

[Publications] Ikeda,M.: "A vacuolar inorganic H^+-pyrophosphatase in Acetabularia acetabulum : Partial purification, characterization and molecular cloning." Journal of Experimental Biology. 50. 139-140 (1998)

[Publications] Sasaki,T.: "Development of vacuoles and vacuolar ATPase activity under extremely high-CO_2 conditions in Chlorococcum littorale cells." Plant Biology. 1(印刷中). (1999)

[Publications] Ueoka-Nakanishi,H.: "Properties and molecular cloning of Ca^<2+>/H^+ antiporter in the vacuolar membrane of mung bean." European Journal of Biochemistry. 印刷中. (1999)

[Publications] Maeshima,M.: "Cycloprodigiosin uncouples H^+-pyrophosphatase of plant vacuolar membrane in the presence of chloride ion." Plant & Cell Physiology. 印刷中. (1999)

[Publications] Nakanishi,Y.: "Molecular cloning and sequencing of the cDNA for vacuolar H^+-pyrophosphatase from Chara corallina." Biochimica et Biophysica Acta. 印刷中. (1999)

[Publications] 前島正義: "細胞工学 別冊 「植物のタンパク質実験」" 液胞膜H^+-ピロホスファターゼの精製, 131-135 (1998)

前島正義名古屋大学, 農学部, 助教授 (80181577)

[Publications] 前島正義: "細胞工学別冊「植物のタンパク質実験」" 液胞膜H^+-ピロホスファターゼの精製, 131-135 (1998)