Calibration and standardisation of synchrotron radiation and conventional circular dichroism spectrometers. Part 2: Factors affecting magnitude and wavelength
Calibration and standardisation of synchrotron radiation and conventional circular dichroism spectrometers. Part 2: Factors affecting magnitude and wavelength
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DOI：
10.1155/2005/263649
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发表时间：
2005-01-01
影响因子：
--
通讯作者：
Wallace, BA
中科院分区：
文献类型：
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作者：
Miles, AJ;Wien, F;Wallace, BA
圆二色光谱是结构生物学中的一种重要工具，特别是用于蛋白质二级结构的分析。同步辐射圆二色性（SRCD）光谱是该技术的修改版本，其使用来自同步加速器源的强光来使数据收集到比传统圆二色性（cCD）仪器可能的波长低得多的波长。有必要对cCD和SRCD仪器之间的校准方法进行标准化，以确保一致性和使用通用参考数据库进行经验二级结构分析的能力。在先前的研究（Spectroscopy 17（2003），653-661）中，我们比较了几种cCD和SRCD仪器上的旋光度测量，同时保持其他实验因素恒定。在这项研究中，其他实验参数，有助于光谱幅度，如细胞路径长度和蛋白质浓度的测定，进行检查。此外，仪器之间的波长校准变化的程度及其对二级结构计算的影响进行了研究。因此，本文件为CD数据计量中的“良好做法”提供了额外的实际指导。
Circular dichroism (CD) spectroscopy is an important tool in structural biology, especially for protein secondary structure analyses. Synchrotron radiation circular dichroism (SRCD) spectroscopy is a modified version of the technique that uses the intense light from a synchrotron source to enable the collection of data to much lower wavelengths than possible on conventional circular dichroism (cCD) instruments. There is a need for standardization of calibration methods amongst and between cCD and SRCD instruments to ensure consistency and the ability to use common reference databases for empirical secondary structural analyses. In a previous study (Spectroscopy 17 (2003), 653-661), we compared optical rotation measurements on several cCD and SRCD instruments, whilst holding constant other experimental factors. In this study, other experimental parameters which contribute to the spectral magnitude, such as cell pathlength and protein concentration determinations, are examined. In addition, the extent of wavelength calibration variations between instruments and their effects on secondary structure calculations have been examined. Hence, this paper provides additional practical guidance for "good practice" in the measurement of CD data.