Unity and diversity of ion transport mechanisms and regulation of Na+/H+ antiporters

离子转运机制的统一性和多样性以及Na /H反向转运蛋白的调节

基本信息

批准号：
13142207
负责人：
KANAZAWA Hiroshi
金额：
$ 54.21万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13142207/
关键词：
intracellular ion homeostasis Na+H+ exchangers pH regulation salinity resistance active transporters molecular structure of membrane proteins intracellular localization of membrane proteins molecular biological approach 細胞内イオン恒常性膜蛋白質

项目摘要

Regulation of intracellular pH, Na^+ concentration, and osmolality are the most important factors for cell survival. These regulations are performed mainly by Na^+/H^+ exchangers named NhaA or NHE on the cytoplasmic as well as endocytic membranes. In this project we aimed to elucidate molecular structure-function relationship and functional regulation of these antiporters including intracellular localization of these molecules for these molecules from bacteria, yeast and mammalian cells. As a result, following achievement was performed. (1) For H pylori NhaA membrane integral domains essential for the ion transport were identified. Further the residues important or essential for Na^+ and H^+ binding were estimated and their hydrophobic environment within the membranes were elucidated. A new detection system of conformational change of NhaA during the ion transport was set up. (2) For yeast Nhalp essential or important residues for the ion transport were identified. Function of the hydrophilic C terminal half domain which seems to be similar to the mammalian NHE were analyzed. A membrane jaxta-positional 16 amino acid residues and its flanking 38 amino acid residues were found to be essential for destination of Nhalp to the cytoplasmic membrane, and binding Cos3p, a noble protein capable enhancing the ion exchange, respectively. (3) For mammalian NHE, we identified two new isoforms, NHE8 and 9. The isoforms NHE 6 to 9 were found to be in the membranes of various endocytic vesicles and function as K+/H+ antipoter rather than Na^+/H^+ antiporter, leading to regulate pH within endocytic vesicles. CHP capable binding NHE1-5 found by us was found to bind other proteins, DRAK2 (apoptotic protein kinase) and KIFIB□ (Kinesin isoform). The present findings will contribute to understand the function and structure relation ship and the regulations of these proteins as well as for elucidating diversity and unity of Na+/H+ antoporters among various organs and species.

细胞内pH、Na^+浓度和渗透压的调节是细胞生存的最重要因素，这些调节主要由细胞质和内吞膜上名为NhaA或NHE的Na^+/H^+交换器执行。在该项目中，我们旨在阐明这些反向转运蛋白的分子结构-功能关系和功能调节，包括这些来自细菌、酵母和哺乳动物细胞的分子的细胞内定位。进一步鉴定了对Na^+和H^+结合重要或必需的残基，并阐明了它们在膜内的疏水环境。一种新的NhaA构象变化检测系统。 (2)对酵母Nhalp的离子转运必需或重要的残基进行了分析，该残基似乎与哺乳动物NHE相似。发现膜 jaxta 位置的 16 个氨基酸残基及其侧翼 38 个氨基酸残基对于 Nhalp 到达细胞质膜以及结合 Cos3p（一种能够增强离子交换的高贵蛋白）至关重要 (3) 对于哺乳动物。 NHE，我们鉴定了两种新的同工型，NHE8 和 9。发现同工型 NHE 6 至 9 存在于各种内吞囊泡的膜中并且作为 K+/H+ 反向转运蛋白而不是 Na^+/H^+ 反向转运蛋白，导致调节内吞囊泡内的 pH 值，我们发现能够结合 NHE1-5 的 CHP 也可以结合其他蛋白质 DRAK2（凋亡蛋白激酶）。本研究结果将有助于理解这些蛋白质的功能和结构关系以及调控，并阐明蛋白质的多样性和统一性。各种器官和物种之间的 Na+/H+ 正向转运蛋白。

项目成果

期刊论文数量（120）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Norihiro Nakamura, Y, Miyake, M.Matsushita, S.Tnaka, H.Inoue, H.Kanazawa: "KIFIBb2, capable of interacting with CHP, is localized to Synaptic vesicles"J. Biochem.. 132. 483-491 (2002)

Norihiro Nakamura、Y、Miyake、M.Matsushita、S.Tnaka、H.Inoue、H.Kanazawa：“KIFIBb2 能够与 CHP 相互作用，定位于突触小泡”J.

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

The murine genome contains one functional gene and two pseudogenes coding for the 16 kDa proteolipid subunit of vacuolar H+-ATPase

小鼠基因组包含 1 个功能基因和 2 个假基因，编码液泡 H -ATP 酶的 16 kDa 蛋白脂质亚基

DOI：
发表时间：
2001
期刊：
Gene 273
影响因子：
0
作者：
Keiko Hayami;Takato Noumi;Hiroki Inoue;Ge-Hong Sun-Wada;Takao Yoshimizu;Hiroshi Kanazawa
通讯作者：
Hiroshi Kanazawa

KIF1Bb2, capable of interacting with CHP, is localized ti synaptic vesicles

KIF1Bb2 能够与 CHP 相互作用，位于突触小泡中

DOI：
发表时间：
2002
期刊：
J.Biochem. 132
影响因子：
0
作者：
N.Nakamura;Y.
通讯作者：
Y.

Hiroki Inoue, Yutaka Nakamura, Mana Nagita, Tomoyo Takai, Miho Masuda, Norihio Nakamura, Hiroshi Kanazawa: "Calcineurin homologous protein isoform 2 (CHP2),Na^+/H^+ exchangers-binding protein, is expressed in intestinal epithelium"Biol.Pharm.Bull.. 26. 14

Hiroki Inoue、Yutaka Nakamura、Mana Nagita、Tomoyo Takai、Miho Masuda、Norihio Nakamura、Hiroshi Kanazawa：“钙调磷酸酶同源蛋白亚型 2 (CHP2)，Na^ /H^ 交换器结合蛋白，在肠上皮中表达”Biol.Pharm