Coherent two-dimensional infrared spectroscopy: Quantitative analysis of protein secondary structure in solution
Coherent two-dimensional infrared spectroscopy: Quantitative analysis of protein secondary structure in solution
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DOI：
10.1039/c2an16031e
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发表时间：
2012-01-01
期刊：
影响因子：
4.2
通讯作者：
Tokmakoff, Andrei
中科院分区：
文献类型：
--
作者：
Baiz, Carlos R.;Peng, Chunte Sam;Tokmakoff, Andrei
我们提出了一种方法来定量测定球状蛋白质的二级结构组成，使用相干二维红外（2 R）光谱骨架酰胺I振动（1550-1720 cm（-1））。16个已知晶体结构的蛋白质被用来构建一个2D光谱库，并通过测量的二维光谱的奇异值分解（SVD）来确定α-螺旋，β-折叠和未分配的构象中的残基的分数。通过从集合中去除每种单独的蛋白质并将从2 μ g中提取的组成与从晶体结构中确定的组成进行比较，对该方法进行基准测试。为了突出从2S光谱中提取的增加的结构内容，还使用文库中蛋白质的常规红外吸收进行了类似的分析。
We present a method to quantitatively determine the secondary structure composition of globular proteins using coherent two-dimensional infrared (2DIR) spectroscopy of backbone amide I vibrations (1550-1720 cm(-1)). Sixteen proteins with known crystal structures were used to construct a library of 2DIR spectra, and the fraction of residues in alpha-helix, beta-sheet, and unassigned conformations was determined by singular value decomposition (SVD) of the measured two-dimensional spectra. The method was benchmarked by removing each individual protein from the set and comparing the composition extracted from 2DIR against the composition determined from the crystal structures. To highlight the increased structural content extracted from 2DIR spectra a similar analysis was also carried out using conventional infrared absorption of the proteins in the library.