Unity and diversity in active transport in cell membrans

细胞膜主动运输的统一性和多样性

基本信息

批准号：
09044310
负责人：
TSUCHIYA Tomofusa
金额：
$ 4.1万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09044310/
关键词：
Active transport Structure and function Unity and diversity Symporter Antiporter

项目摘要

It is important to analyze many transport systems for the investigation of unity and diversity in structure, function and mechanism in active transport proteins. Fortunately, we have found and investigated many active transport systems in bacterial cells so far. In the past two years, we have obtained the following results.1) We analyzed many NaィイD1+ィエD1-coupled transport systems, mainly NaィイD1+ィエD1/melibiose symporter and NaィイD1+ィエD1/serine symporter of Escherichia coli and some other bacteria. We characterized the symporters from biochemical view point and genetic view point. We established methods for over production, large-scale purification and reconstitution of the NaィイD1+ィエD1/melibiose symporter protein and NaィイD1+ィエD1/serine symporter protein. This opened up a way for further analysis in structure, function and mechanism an active transport proteins.2) We identified regions and amino acid residues important or necessary for ion-coupling or substrate recognition in the NaィイD1+ィエD1/melibiose symporter protein through mutant analysis and site-directed mutagenesis. Homology search and motif analysis were also useful for such analysis.3) We found new transport systems and cloned their genes in some bacteria. Biochemical and genetic analyses further revealed unity and diversity in transport proteins.Collaborations with Prof. Wilson and Prof. Konings were very valuable foe this project.

分析多种转运系统对于研究主动转运蛋白结构、功能和机制的统一性和多样性具有重要意义。在过去的两年里，我们已经发现并研究了细菌细胞中的多种主动转运系统。得到以下结果。 1)我们分析了许多NaiD1+D1偶联转运系统，主要是NaiD1+D1/蜜二糖同向转运体和NaiD1+D1/丝氨酸同向转运体我们从生化角度和遗传学角度对大肠杆菌和其他一些细菌进行了同向转运蛋白的表征，我们建立了 NaiD1+D1/蜜二糖同向转运蛋白和 NaiI D1+D1/丝氨酸同向转运蛋白的过量生产、大规模纯化和重建方法。这为进一步分析主动转运蛋白的结构、功能和机制开辟了道路。2)我们鉴定了重要或必需的区域和氨基酸残基。通过突变体分析和定点诱变对 NaiID1+IeD1/蜜二糖同向转运蛋白进行离子偶联或底物识别也可用于此类分析。3) 我们发现了新的转运系统并在一些细菌中克隆了它们的基因。生化和遗传分析进一步揭示了转运蛋白的统一性和多样性。与 Wilson 教授和 Konings 教授的合作对于这个项目非常有价值。