Wien Frank, Geinguenaud Frédéric, Grange Wilfried, Arluison Véronique
Synchrotron SOLEIL, L'Orme des Merisiers Saint Aubin, Gif-sur-Yvette, France.
Plateforme CNanoMat, UFR SMBH, Université Paris 13, Bobigny, France.
Methods Mol Biol. 2021;2209:87-108. doi: 10.1007/978-1-0716-0935-4_6.
Fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopies are fast techniques providing important information about the conformation of nucleic acids and proteins. These vibrational and electronic absorption spectroscopies are extremely sensitive to any change in molecular structure. While numerous reviews describe how to analyze DNA structure alone or in the presence of proteins using FTIR and CD, analyses of RNA are scarce. Nevertheless, RNA remodeling proteins are important factors involved in a multitude of roles in the cell. In this chapter, we present applications of synchrotron radiation circular dichroism (SRCD) and FTIR to analyze how proteins may change RNA structure. These include the analysis of RNA melting, or stabilization, of change in helical parameters and base stacking. The effects on the structure of RNA remodeling proteins are also presented.
傅里叶变换红外光谱(FTIR)和圆二色光谱(CD)是快速技术,可提供有关核酸和蛋白质构象的重要信息。这些振动光谱和电子吸收光谱对分子结构的任何变化都极为敏感。虽然众多综述描述了如何单独使用FTIR和CD分析DNA结构,或在蛋白质存在的情况下分析DNA结构,但对RNA的分析却很少。然而,RNA重塑蛋白是细胞中多种作用的重要因素。在本章中,我们介绍同步辐射圆二色光谱(SRCD)和FTIR在分析蛋白质如何改变RNA结构方面的应用。这些应用包括对RNA解链、稳定化、螺旋参数变化和碱基堆积变化的分析。同时还介绍了对RNA重塑蛋白结构的影响。