Protein structure analysis by robust electron spin resonance at the single molecule level.

在单分子水平上通过强大的电子自旋共振进行蛋白质结构分析。

• 批准号: 392072074
• 负责人: Professor Dr. Friedemann Reinhard
• 金额: --
• 依托单位: Institut für Physik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/392072074?language=en
• 关键词: Protein; structure; analysis; robust; electron

The project aims to demonstrate analysis of structure and dynamics of a protein by single-molecule electron spin spectroscopy. This ambitious goal has come within reach by recent joint work of the applicants: ESR detection of a single protein, labeled by a stable organic electron spin, using a nearby Nitrogen-Vacancy (NV) color center in diamond as a sensor [F. Shi, et al., Science 347, 1135 (2015)]. The project will overcome two major problems encountered in this proof-of-principle experiment: low stability of the spin labels involved and inefficient detection. This shall be accomplished by two approaches: resonant single-photon and two-photon excitation of the NV sensor at cryogenic temperature; use of more resilient labels and nano-photonic structures for readout enhancement at room temperature.The project will equally develop protocols for preparation of single site-specifically labeled proteins on the sensor surface and structure determination by multidimensional spectroscopy. It will finally combine all these developments to enable stable and efficient spectroscopy of single molecules by electron spin resonance, in particular three-dimensional structure analysis of multiply labeled proteins.

该项目旨在通过单分子电子自旋光谱法演示蛋白质的结构和动力学分析。通过申请人最近的联合工作，这一雄心勃勃的目标已经实现：ESR检测单个蛋白质，由稳定的有机电子自旋标记，使用金刚石中附近的氮空位（NV）色心作为传感器[F. Shi等人，Science 347，1135（2015）]。该项目将克服在该原理验证实验中遇到的两个主要问题：所涉及的自旋标签的低稳定性和检测效率低下。这将通过两种方法来实现：在低温下NV传感器的共振单光子和双光子激发;在室温下使用更具弹性的标签和纳米光子结构来增强读出。该项目将同样开发用于在传感器表面制备单位点特异性标记蛋白质和通过多维光谱确定结构的协议。它最终将联合收割机结合所有这些发展，通过电子自旋共振实现单分子的稳定和高效光谱分析，特别是多重标记蛋白质的三维结构分析。