Del Villar-Guerra Rafael, Gray Robert D, Chaires Jonathan B
James Graham Brown Cancer Center, University of Louisville, Louisville, Kentucky.
Curr Protoc Nucleic Acid Chem. 2017 Mar 2;68:17.8.1-17.8.16. doi: 10.1002/cpnc.23.
Circular dichroism (CD) is a phenomenon that arises from the differential absorption of left- and right-handed circularly polarized light, and may be seen with optically active molecules. CD spectroscopy provides useful spectral signatures for biological macromolecules in solution, and provides low-resolution structural information about macromolecular conformation. CD spectroscopy is particularly useful for monitoring conformational changes in macromolecules upon environmental perturbations. G-quadruplex structures show unique CD spectral signatures, and CD is an important tool for characterizing their formation and global structure. This protocol offers step-by-step methods for determining reliable and reproducible CD spectra of quadruplex structures and normalizing the spectra for presentation. CD spectra properly normalized with respect to quadruplex concentration and path length are required to facilitate accurate comparison of results among laboratories. The standard operating procedures proposed are recommended to make such comparison accurate and informative. © 2017 by John Wiley & Sons, Inc.
圆二色性(CD)是一种由左旋和右旋圆偏振光的差异吸收引起的现象,在光学活性分子中可见。CD光谱为溶液中的生物大分子提供了有用的光谱特征,并提供了有关大分子构象的低分辨率结构信息。CD光谱对于监测环境扰动下大分子的构象变化特别有用。G-四链体结构显示出独特的CD光谱特征,并且CD是表征其形成和整体结构的重要工具。本方案提供了逐步方法,用于确定四链体结构可靠且可重复的CD光谱,并对光谱进行归一化以用于展示。为便于实验室间结果的准确比较,需要根据四链体浓度和光程对CD光谱进行适当归一化。建议采用所提出的标准操作程序,以使这种比较准确且信息丰富。© 2017约翰威立父子公司版权所有