Structure and mechanism in cation-coupled transport systems

阳离子耦合传输系统的结构和机制

基本信息

  • 批准号:
    04044122
  • 负责人:
  • 金额:
    $ 3.9万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for international Scientific Research
  • 财政年份:
    1992
  • 资助国家:
    日本
  • 起止时间:
    1992 至 1993
  • 项目状态:
    已结题

项目摘要

Cation/substrate symport is a major mechanism of active solute transport in cell membranes. In many cation/substrate symport systems, Na^+ or H^+ is utilized as the coupling cation. Namely, the driving force for the symport is an electrochemical potential of Na^+ or H^+. An electrochemical potential of Na^+ is established by Na^+/H^+ antiporter. In this study, we investigated structure of the transport proteins and mechanism of transport in the symporter and the antiporter.The melibiose transport system of Escherichia coli and Salmonella Typhimurius utilizes either Na^+, H^+ or Li^+ as the coupling cation for symport depending on substrate transported. We isolated many types of mutant which showed altered cation specificity, altered substrate specificity, altered activity or altered temperature sensitivity. We cloned the mutant types melB gene, identified substitution of nucleotide and identified substituted amino acid residues in the melibiose transport protein. We found many Pro to Ser replacements. Then we constructed many mutant types of the melibiose transport protein by site-directed mutagenesis. Properties of the mutated melibiose transport proteins were analyzed. Thus, we revealed structure-function relationship and role of amino acid residues in the melibiose transport protein.Function of the melibiose transport system is regulated by phospho-transferase system. We identified amino acid residues and domain in the melibiose transporter, which are involved in the interaction with a regulatory protein.We also characterized Na^+/H^+ antiporter, and cloned and sequenced nhaA gene encoding the antiporter. Thus, we obtained information about structure-function relationship in the Na^+/H^+ antiporter.
阳离子/底物同调是细胞膜内溶质主动转运的主要机制。在许多阳离子/衬底共模体系中,Na^+或H^+被用作耦合阳离子。也就是说,对称的驱动力是Na^+或H^+的电化学电位。Na^+的电化学电位由Na^+/H^+反转运体确定。在本研究中,我们研究了转运蛋白在正转运蛋白和反转运蛋白中的结构和转运机制。大肠杆菌和鼠伤寒沙门氏菌的糖二糖转运系统利用Na^+、H^+或Li^+作为同调的偶联阳离子,这取决于所转运的底物。我们分离出许多类型的突变体,它们表现出阳离子特异性改变,底物特异性改变,活性改变或温度敏感性改变。我们克隆了突变型melB基因,鉴定了甲基二糖转运蛋白中核苷酸的替换和氨基酸的替换残基。我们发现了许多Pro到Ser的替代品。然后,我们通过位点定向诱变构建了多种突变型的糖二糖转运蛋白。分析了突变的糖二糖转运蛋白的特性。因此,我们揭示了氨基酸残基在糖二糖转运蛋白中的结构-功能关系和作用。二糖转运系统的功能受磷酸转移酶系统的调控。我们鉴定了二糖转运体中的氨基酸残基和结构域,这些氨基酸残基和结构域参与了与调节蛋白的相互作用。我们还鉴定了Na^+/H^+反转运蛋白,并克隆了编码该反转运蛋白的nhaA基因并对其进行了测序。由此,我们获得了Na^+/H^+反转运子的结构-功能关系信息。

项目成果

期刊论文数量(15)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Masayuki Kuroda: "Resistance of the melibiose carrier to inhibition by the phosphotransferase system due to substitutions of amino acid residues in the carrier of Salmonella typhimurium" Journal of Biological Chemistry. 267. 18336-18341 (1992)
Masayuki Kuroda:“由于鼠伤寒沙门氏菌载体中氨基酸残基的取代,蜜二糖载体对磷酸转移酶系统的抑制具有抵抗力”《生物化学杂志》。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
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  • 通讯作者:
Noriko Okazaki: "Characterization of the lactose transport system in Citrobacter freundii" Biological Pharmaceutical Bulletin. (印刷中). (1994)
Noriko Okazaki:“弗氏柠檬酸杆菌乳糖转运系统的表征”生物制药通报(1994 年)。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
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  • 通讯作者:
Kei Inaba: "Lithium toxicity and Na^+/H^+ antiporter in Escherichia coli" Biological Pharmaceutical Bulletin. (印刷中). (1994)
Kei Inaba:“大肠杆菌中的锂毒性和 Na^+/H^+ 反向转运蛋白”生物制药通报(1994 年)。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Masayuki Kuroda: "Resistance of the melibiose carrier to inhibition by the phosphotransferase system due to substitutions of amino acid residues in the carrier of Salmonella typhimurium" The Journal of Biological Chemistry. 267. 18336-18341 (1992)
Masayuki Kuroda:“由于鼠伤寒沙门氏菌载体中氨基酸残基的取代,蜜二糖载体对磷酸转移酶系统抑制的抵抗力”《生物化学杂志》。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Masayuki Kuroda: "Resistance of the melibiose carrier to inhibition by the phosph-transferase system due to substitutions of amino acid residues in the carrier of Salmonella typhimurium" Journal of Biological Chemistry. 267. 18336-18341 (1992)
Masayuki Kuroda:“由于鼠伤寒沙门氏菌载体中氨基酸残基的取代,蜜二糖载体对磷酸转移酶系统抑制的抵抗力”《生物化学杂志》。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
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TSUCHIYA Tomofusa其他文献

TSUCHIYA Tomofusa的其他文献

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{{ truncateString('TSUCHIYA Tomofusa', 18)}}的其他基金

Analysis of multidrug resistance systems in multidrug resistant bacteria and development of drugs effective on the resistant bacteria
多重耐药菌的多重耐药系统分析及针对耐药菌有效药物的开发
  • 批准号:
    20590120
  • 财政年份:
    2008
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Systematic analysis of multidrug efflux pumps in multidrug resistant bacteria and development of pump inhibitor
多重耐药菌多药外排泵的系统分析及泵抑制剂的开发
  • 批准号:
    16390131
  • 财政年份:
    2004
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Tne initiator of chromosomal DNA replication in E. coli.
大肠杆菌中染色体 DNA 复制的引发剂。
  • 批准号:
    11470486
  • 财政年份:
    1999
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Unity and diversity in active transport in cell membrans
细胞膜主动运输的统一性和多样性
  • 批准号:
    09044310
  • 财政年份:
    1997
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
Structure and function of drug- and ion-extrusion systems in bacteria
细菌中药物和离子挤出系统的结构和功能
  • 批准号:
    09672230
  • 财政年份:
    1997
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Characteristics of ion tranport systems in Vibrio parahaemolyticus and Staphylococcus aureus
副溶血性弧菌和金黄色葡萄球菌离子传输系统的特点
  • 批准号:
    07672358
  • 财政年份:
    1995
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Molecular mechanism of energy coupling in active transport
主动运输中能量耦合的分子机制
  • 批准号:
    06044153
  • 财政年份:
    1994
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
Analysis of factors involved in serine-sensitivity in bacterial cells
细菌细胞丝氨酸敏感性相关因素分析
  • 批准号:
    04671351
  • 财政年份:
    1992
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
Structure and mechanism in active transport systems for sugars, amino acids and ions.
糖、氨基酸和离子主动运输系统的结构和机制。
  • 批准号:
    63571041
  • 财政年份:
    1988
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
Alterations in the primary structure of the active transport carrier and changes in the function
主动转运载体一级结构的改变和功能的变化
  • 批准号:
    61580145
  • 财政年份:
    1986
  • 资助金额:
    $ 3.9万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
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