Structural and Functional Analysis of Bacterial Formate Channels

细菌甲酸通道的结构和功能分析

• 批准号: 197321781
• 负责人: Professor Dr. Oliver Einsle
• 金额: --
• 依托单位: Institut für Biochemie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/197321781?language=en
• 关键词: Structural; Functional; Analysis; Bacterial; Formate

Formate constitutes a major metabolite in bacteria and plays a key role as electron donor in various respiratory pathways. The transport of formate across biological membranes is mediated by the formate channel FocA, a member of the FNT family of membrane transporters. Our three-dimensional structure of the protein from Salmonella typhimurium at low pH showed a concerted and cooperative set of conformational changes of the N-terminus that form the basis for a functional switch from the export of the formate anion by facilitated diffusion at high external pH to secondary active formate/H+ import at a pH below 6.8. This corresponds to a metabolic shift from cytoplasmic formate production by pyruvate:formate lyase (PFL) and periplasmic, respiratory formate oxidation to cytoplasmic formate disproportionation at the formate:hydrogen lyase complex (FHL). We propose to continue and extend our work on FocA to include an electrophysiological characterization of the formate channel and its dependency on the proton motive force in lipid bilayers. We will further analyse the mechanism of pH sensing in FocA by site-directed mutagenesis and investigate the subcellular localization of the protein and its interaction with physiological partners such as PFL or the FHL complex.

甲酸盐是细菌的主要代谢产物，在多种呼吸途径中作为电子供体发挥关键作用。甲酸盐跨生物膜的转运由甲酸盐通道福卡介导，该通道是膜转运蛋白FNT家族的成员。我们的三维结构的蛋白质从鼠伤寒沙门氏菌在低pH值显示了一个协调和合作的N-末端的构象变化，形成了一个功能开关的基础上，从出口的甲酸阴离子的促进扩散在高外部pH值的二级活性甲酸/H+进口在pH值低于6.8。这对应于从丙酮酸：甲酸裂解酶（PFL）和周质呼吸性甲酸氧化的细胞质甲酸生产到甲酸：氢裂解酶复合物（FHL）的细胞质甲酸分解的代谢转变。我们建议继续和扩展我们的工作福卡，包括甲酸通道的电生理特性和它的依赖性的质子动力在脂质双层。我们将通过定点突变进一步分析福卡的pH传感机制，并研究蛋白质的亚细胞定位及其与生理伙伴如PFL或FHL复合物的相互作用。

项目成果

The tricky task of nitrate/nitrite antiport.

硝酸盐/亚硝酸盐逆向转运的棘手任务

• DOI: 10.1002/anie.201305421
• 发表时间: 2013
• 期刊: Angewandte Chemie
• 作者: Andrade SLA ;Einsle O
• 通讯作者: Einsle O

Structural and Functional Studies of NirC from Salmonella typhimurium.

鼠伤寒沙门氏菌 NirC 的结构和功能研究

• DOI: 10.1016/bs.mie.2014.12.034
• 发表时间: 2015
• 期刊: Methods in enzymology
• 作者: Rycovska-Blume A;Andrade SLA;Fendler K ;Einsle O
• 通讯作者: Einsle O