Dynamics and Function of Spin Labelled ABC Transport System Studied by Multi-Frequency EPR

多频EPR研究自旋标记ABC转运系统的动力学和功能

• 批准号: 137912997
• 负责人: Professor Dr. Heinz-Jürgen Steinhoff
• 金额: --
• 依托单位: Arbeitsgruppe Makromolekülstruktur
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/137912997?language=en
• 关键词: Dynamics; Function; Spin; Labelled; ABC

The current transport model of ABC transporters is essentially based on the biochemical, biophysical and structural investigations of the maltose transporter MalFGK2 of E. coli/Salmonella. Due to some unique features of this system it is questionable whether this transport model holds for all ABC transporters. Here, the histidine ABC transporter from Salmonella typhimurium, HisJ/LAO-HisQMP2, and of the ECF (“energy-coupling factor”) transporter family, the biotin transporter BioMNY are regarded as complementary model systems for ABC transporters. Based on our recent results we will focus now on the interaction between the substrate binding protein HisJ and HisQMP2 and on substrate and nucleotide induced conformational changes of ECF transporters. The functionally related conformational changes of site-directed spin labeled variants of these transporters will be followed during the transport cycle by multi-frequency cw and pulse EPR spectroscopy with the aim to unravel the transport mechanism(s) on molecular level. In addition we intend to develop and apply new methodological approaches in the field of EPR spectroscopy including MD simulation based EPR spectra calculations.

目前ABC转运蛋白的转运模型基本上是基于对大肠杆菌麦芽糖转运蛋白MalFGK 2的生化、生物物理和结构研究。大肠杆菌/沙门氏菌。由于这个系统的一些独特的功能，这是值得怀疑的运输模式是否适用于所有ABC运输。在此，来自鼠伤寒沙门氏菌的组氨酸ABC转运蛋白HisJ/LAO-HisQMP 2和ECF（“能量偶联因子”）转运蛋白家族的生物素转运蛋白BioMNY被视为ABC转运蛋白的互补模型系统。基于我们最近的研究结果，我们现在将重点放在底物结合蛋白HisJ和HisQMP 2之间的相互作用和底物和核苷酸诱导的ECF转运蛋白的构象变化。这些转运蛋白的定点自旋标记变体的功能相关构象变化将在转运循环期间通过多频cw和脉冲EPR光谱进行跟踪，目的是在分子水平上阐明转运机制。此外，我们打算开发和应用新的方法学方法在EPR光谱领域，包括基于分子动力学模拟EPR谱计算。

项目成果

Conformational changes of the histidine ATP-binding cassette transporter studied by double electron-electron resonance spectroscopy.

双电子-电子共振光谱研究组氨酸 ATP 结合盒转运蛋白的构象变化

• DOI: 10.1016/j.bbamem.2014.02.010
• 发表时间: 2014
• 期刊: Biochimica et biophysica acta
• 作者: Sippach M;Weidlich D;Klose D;Abé C;Klare J;Schneider E;Steinhoff H-J
• 通讯作者: Steinhoff H-J