DNP enhanced MAS solid-state NMR of biological systems in their native context

生物系统在其原生环境中的 DNP 增强 MAS 固态 NMR

• 批准号: 226561002
• 负责人: Professor Dr. Bernd Reif
• 金额: --
• 依托单位: Professur für Festkörper-NMR-Spektroskopie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/226561002?language=en
• 关键词: DNP; enhanced; MAS; solid; state

We propose to use DNP (Dynamic Nuclear Polarization) MAS solid-state NMR to determine the structure of membrane proteins and amyloid aggregates in their native cellular environment without the need for purification and reconstitution. The method will be applied to membrane proteins that we characterized previously employing standard NMR methodologies. In particular, we will focus on the membrane protein Mistic, a membrane protein chaperone, and EmrE, a multidrug resistance transporter. Furthermore, we will investigate the Alzheimer’s disease ß-amyloid peptide and characterize its structure in the context of neuronal cells. DNP is essential in all these projects as the available amount of isotopically enriched material, which can be packed into a MAS rotor, and its concentration is typically too little to enable a proper structural characterization using standard solution-state or solid-state NMR spectroscopy methods.

我们建议使用DNP（动态核极化）MAS固态NMR来确定膜蛋白和淀粉样蛋白在其天然细胞环境中的结构，而无需纯化和重建。该方法将应用于我们使用标准NMR方法的先前表征的膜蛋白。特别是，我们将重点放在膜蛋白薄膜（一种膜蛋白伴侣）和多药耐药转运蛋白EMRE上。此外，我们将研究阿尔茨海默氏病β-淀粉样蛋白肽，并在神经元细胞的背景下表征其结构。 DNP在所有这些项目中都是必不可少的，因为可用的同位素富集材料可以包装成MAS转子，并且其浓度通常太少，无法使用标准溶液状态或固态NMR光谱法实现适当的结构表征。

项目成果

The mechanism of denaturation and the unfolded state of the α-helical membrane-associated protein Mistic.

α螺旋膜相关蛋白Mistic的变性机制和展开状态

• DOI: 10.1021/ja408644f
• 发表时间: 2013
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Jacso T;Bardiaux B;Broecker J;Fiedler S;Barwinkel T;Mainz A;Fink U;Vargas C;Oschkinat H;Keller S;Reif B
• 通讯作者: Reif B