STRUCTURE AND FUNCTION OF ANIMAL ION TRANSPORTER THAT EXPRESS IN BACTERIA

细菌中表达的动物离子转运蛋白的结构和功能

• 批准号: 10672037
• 负责人: NAKAMURA Tatsunosuke
• 金额: $ 1.6万
• 依托单位: CHIBA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672037/
• 关键词: TRP; cation channel; Escherichia coli; capsaicin receptor; K^+ channel; structure and function; mammalian cultured cells; イオン選択性; チャネル; Na^+排出; K^+輸送系; トランスポーター; antiporter; イオン輸送系; 蛋白発現; クロストーク; ポア構造

TRP (transient receptor rotential) homologue of mouse function as cation channel that passes Ca^<2+> and Na^+. When TRP homologue gene was transformed in Escherichia coli mutant strain TO114 which lacks all Na^+/H^+ antiporters, this strain became sensitive to Na^+ suggesting that it functions in E.coli membrane. We detected His tagged TRP homologue expressed in membrane fraction of E.coli. TRP probably has common structure with VR1, capsaicin receptor, but those structures are not known. TRP/VR1 were suggested to have homology with Shaker type K^+ channels with 6 transmembrane regions, though there are no experimental reports. We started experiment for TRP structure using bacterial system and got some results. We are doing further experiment using mammalian cultured cells to confirm the TRP structure that got from bacterial system.

小鼠TRP（瞬时受体电位）同源物作为阳离子通道发挥作用，通过Ca^2+和Na^+。当TRP同源基因转化到缺乏所有Na^+/H^+反向转运蛋白的大肠杆菌突变株TO 114中时，该菌株变得对Na^+敏感，这表明它在大肠杆菌膜中起作用。我们检测到His标记的TRP同源物在大肠杆菌的膜部分中表达。TRP可能与辣椒素受体VR 1具有共同的结构，但这些结构尚不清楚。TRP/VR 1被认为与具有6个跨膜区的Shaker型K^+通道具有同源性，但还没有实验报道。我们利用细菌系统进行了TRP结构的实验，并得到了一些结果。我们正在用哺乳动物培养细胞做进一步的实验，以证实从细菌系统中得到的TRP结构。

项目成果

A.Hamada: "The Arabidopsis HKT1 Gene Homolog Mediates Inward Na^+ Currents in Xenopus laevis Oocytes and Na^+ Uptake in Saccharomyces cerevisiae."Plant Physiology. 125. 437-446 (2001)

A.Hamada：“拟南芥 HKT1 基因同源物介导非洲爪蟾卵母细胞的内向 Na^ 电流和酿酒酵母的 Na^ 吸收。”植物生理学。

Y.Nakamaru, Y.Takahashi, T.Unemoto, T.Nakamura.: "Mechanosensitive channel (MscL) functions to alleviate the cell lysis of marine bacterium, Vibrio alginolyt."FEBS Letters. 444. 170-172 (1999)

Y.Nakamaru、Y.Takahashi、T.Unemoto、T.Nakamura.：“机械敏感通道 (MscL) 的功能是减轻海洋细菌溶藻弧菌的细胞裂解。”FEBS 快报。

N.Uozumi: "The Arabidopsis HKT1 Gene Homolog Mediates Inward Na^+ Currents in Xenopus laevis Oocytes and Na^+ Uptake in Saccharomyces cerevisiae."Plant Physiology. 122. 1249-1260 (2000)

N.Uozumi：“拟南芥 HKT1 基因同源物介导非洲爪蟾卵母细胞的内向 Na^ 电流和酿酒酵母的 Na^ 吸收。”植物生理学。

T.Nakamura, N.Yamamuro, S.Stumpe, T.Unemoto, E.P.Bakker.: "Cloning of the trkAH gene cluster and characterization of the Trk K^+-uptake system of Vibrio alginoyticus."Microbiology. 144. 2281-2289 (1998)

T.Nakamura、N.Yamamuro、S.Stumpe、T.Unemoto、E.P.Bakker.：“藻酸弧菌 trkAH 基因簇的克隆和 Trk K^-摄取系统的表征。”微生物学。

P.Putnoky, A.Kereszt, T.Nakamura, G.Endre, E.Grosskopf, P.Kiss, A.Kondorosi.: "The pha gene cluster of Rhizobium meliloti involved in pH adaptation and symbiosis encodes a novel type of K^+ efflux system."Molecular Microbiology. 28. 1091-1101 (1998)

P.Putnoky、A.Kereszt、T.Nakamura、G.Endre、E.Grosskopf、P.Kiss、A.Kondorosi.：“苜蓿根瘤菌的 pha 基因簇参与 pH 适应和共生，编码一种新型 K^