Joint Study on the Molecular Biology of SA Channels

SA通道分子生物学联合研究

• 批准号: 07044245
• 负责人: SOKABE Masahiro
• 金额: $ 2.5万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07044245/
• 关键词: SA channel; me c-gene; Drosophila; volume regulation; antibody; opider venom; cGMP依存性チャネル; 遺伝子クローニング; 分子生物学; MEC遺伝子; A6細胞

Stretch activated (SA) ion channels are expressed in many types of cells and supposed to play an important role in fundamental cell functions including cell growth, division, morphogenesis, and volume regulation. However, because of the lack of information on their molecular structure, further exploration of SA channels is confronted with difficulties. Very recently a gene encoding an SA channel has been isolated from E.coli, however, its shows a peculiar sequence devoid of any homology with known sequences of ion channels in eucaryotic cels. The aim of the present study was to find SA channel genes from eucaryotic cells and to develop a specific blockers to the SA channel, by which we could explore structure-function and physiological roles of SA channels. We found a candidate gene in MDCK cells of which sequence has a significant homology with mec gene that is supposed to be a putative SA channel gene. We also found, in Drosophila, a gene encoding new type of cGMP gated ion channel that may contribute to mechanotransduction in this species. Expression and functional assay of these genes are in progress. A certain spider venom has been found to include peptides that specifically block the SA channel. We succeeded in purifying some of these peptides with defferent degree of affinity to the SA channel. The structure of a low a affinity one has been sccessfully determined. Determination of a high affinity one is now in progress. We are planning to prepare an affinity column by using the peptide to purify SA channel proteins. We are also going to prepare a fluorescent-conjugated peptide by which the distribution of SA channels on the cell surface could be studied.

拉伸活化的（SA）离子通道在许多类型的细胞中表达，并且应该在基本细胞功能中起重要作用，包括细胞生长，分裂，形态发生和体积调节。但是，由于缺乏有关其分子结构的信息，SA通道的进一步探索面临困难。最近，已经从大肠杆菌中分离出一个编码SA通道的基因，但是它显示出具有与已知的离子通道序列中的任何同源性的奇特序列。本研究的目的是找到来自桉树细胞的SA通道基因，并向SA通道开发特定的阻滞剂，我们可以探索SA通道的结构功能和生理作用。我们在MDCK细胞中发现了一个候选基因，该基因与MEC基因具有重要的同源性，该基因被认为是推定的SA通道基因。我们还发现，在果蝇中，一种编码新型CGMP门控离子通道的基因，可能有助于该物种的机械转导。这些基因的表达和功能测定正在进行中。已经发现某种蜘蛛毒液包括特异性阻断SA通道的肽。我们成功地净化了其中一些与SA通道相关程度的这些肽。低A亲和力的结构已被确定。确定高亲和力的确定正在进行中。我们计划通过使用肽净化SA通道蛋白来准备一个亲和力柱。我们还将准备一个荧光偶联的肽，通过该肽可以研究细胞表面上的SA通道的分布。

项目成果

曽我部正博: "単一チャネルデータの処理と解析法 In パッチクランプ実験技術法(岡田編)pp50-62" 吉岡書店, 181 (1996)

Masahiro Sogabe：“膜片钳实验技术中的单通道数据处理和分析方法（冈田编）第50-62页”吉冈书店，181（1996）

曽我部正博: "パッチクランプ法による単一チャネル電流の記録 In 英語論文セミナー、現代の細胞生物学(馬淵、山本編)pp113-120" 講談社, 168 (1996)

Masahiro Sogabe：“通过膜片钳方法记录单通道电流在英文论文研讨会中，现代细胞生物学（Mabuchi和Yamamoto编辑）第113-120页”讲谈社，168（1996）

曽我部正博: "細胞機能 In「アトラスで学ぶ生理学」(高田編) pp2-13" 丸善, 281 (1996)

曾我部正宏：《用 Atlas 学习生理学》中的细胞功能（高田编辑）第 2-13 页”Maruzen，281（1996）

曽我部正博: "機械受容、容量受容器、In「分子細胞生物学辞典」(村松編)" 東京化学同人(印刷中),

Masahiro Sogabe：“机械感受，电容性感受器，《分子细胞生物学辞典》（村松编）”东京化学同人（出版中），

曽我部正博: "細胞機能. In「アトラスの生理学」(高田編)" 丸善(印刷中),

Masahiro Sogabe：“细胞功能。在“阿特拉斯生理学”（高田编辑）”丸善（出版中），