DETERMINATION OF THE SECONDARY STRUCTURE OF PROTEINS FROM THE RAMAN AMIDE-I BAND - THE REFERENCE INTENSITY PROFILES METHOD
DETERMINATION OF THE SECONDARY STRUCTURE OF PROTEINS FROM THE RAMAN AMIDE-I BAND - THE REFERENCE INTENSITY PROFILES METHOD
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DOI：
10.1002/jrs.1250180411
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发表时间：
1987-06-01
影响因子：
2.5
通讯作者：
ALIX, AJP
中科院分区：
文献类型：
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作者：
BERJOT, M;MARX, J;ALIX, AJP
提出了一种从拉曼光谱中确定蛋白质定量二级结构的简单方法，即参考强度分布法。它使用主链酰胺 I 振动的拉曼光谱强度（范围 1630-1700 cm−1）作为模型，对应于纯二级构象，即 α 螺旋、β 片层和未定义的结构。参考强度分布是通过对大量蛋白质的实验酰胺 I 光谱进行逐步拟合来计算的，这些蛋白质的二级结构从固态 X 射线衍射数据中众所周知。该方法可用于确定任何状态下蛋白质的二级结构。对于溶液中的蛋白质，该方法同时求解二级结构含量和减法系数，从而确定溶剂光谱的贡献。所提出的方法已成功应用于大量蛋白质。对于每一类结构，X 射线数据与拉曼计算结果之间的相关系数接近 0.97，因此可以使用参考强度剖面方法得出二级结构的准确估计。
A simple method for determining the quantitative secondary structure of proteins from their Raman spectra, the reference intensity profiles method, is proposed. This uses as a model the Raman spectral intensities of the backbone amide I vibrations (range 1630–1700 cm−1), corresponding to pure classes of secondary conformations, i.e. α‐helical, β‐sheet and undefined structures. The reference intensity profiles were computed by a step‐by‐step fit of the experimental amide I spectra of a large number of proteins, the secondary structures of which were well known from x‐ray diffraction data in the solid state. The method could be of use in determining the secondary structure of a protein in any state. In the case of a protein in solution, the method solves simultaneously for the secondary structural contents and the subtraction coefficient, thus determining the solvent spectral contribution. The proposed method has been applied with success to a large set of proteins. The correlation coefficient between x‐ray data and the Raman calculated results is close to 0.97 for each class of structure, so accurate estimates of secondary structure may be derived by using the reference intensity profiles method.