Biol Chem. 2013 Oct;394(10):1281-300. doi: 10.1515/hsz-2013-0155.
Site-directed spin labeling (SDSL) in combination with electron paramagnetic resonance (EPR) spectroscopy has emerged as an efficient tool to elucidate the structure and the conformational dynamics of proteins under conditions close to the native state. This review article summarizes the basics as well as the recent progress in SDSL and EPR methods, especially for investigations on protein structure, protein function, and interaction of proteins with other proteins or nucleic acids. Labeling techniques as well as EPR methods are introduced and exemplified with applications to systems that have been studied in the author's laboratory in the past 15 years, headmost the sensory rhodopsin-transducer complex mediating the photophobic response of the halophilic archaeum Natronomonas pharaonis. Further examples underline the application of SDSL EPR spectroscopy to answer specific questions about the system under investigation, such as the nature and influence of interactions of proteins with other proteins or nucleic acids. Finally, it is discussed how SDSL EPR can be combined with other biophysical techniques to combine the strengths of the different methodologies.
定点自旋标记（SDSL）与电子顺磁共振（EPR）光谱学相结合，已成为在接近天然状态的条件下阐明蛋白质结构和构象动力学的有效工具。本文综述了 SDSL 和 EPR 方法的基础和最新进展，特别是在蛋白质结构、蛋白质功能以及蛋白质与其他蛋白质或核酸相互作用的研究方面。本文介绍了标记技术和 EPR 方法，并举例说明了过去 15 年中作者实验室研究过的系统的应用，其中首先是介导嗜盐古菌盐沼甲烷单胞菌光回避反应的感觉视紫红质转导复合物。进一步的例子强调了 SDSL EPR 光谱学在回答关于所研究系统的具体问题中的应用，例如蛋白质与其他蛋白质或核酸相互作用的性质和影响。最后，讨论了如何将 SDSL EPR 与其他生物物理技术相结合，以结合不同方法学的优势。
