UV resonance Raman-selective amide vibrational enhancement: Quantitative methodology for determining protein secondary structure
UV resonance Raman-selective amide vibrational enhancement: Quantitative methodology for determining protein secondary structure
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DOI：
10.1021/bi971160z
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发表时间：
1998-03-03
期刊：
影响因子：
2.9
通讯作者：
Asher, SA
中科院分区：
文献类型：
--
作者：
Chi, ZH;Chen, XG;Asher, SA
我们通过在206.5纳米处激发酰胺π→π*跃迁，直接测定了“平均”纯α - 螺旋、β - 折叠和无规二级结构的酰胺带共振拉曼光谱。拉曼光谱主要由肽主链的酰胺带所主导。我们从13种具有已知X射线晶体结构的蛋白质的酰胺共振拉曼光谱中，凭经验确定了平均纯α - 螺旋、β - 折叠和无规共振拉曼光谱。我们证明，我们可以同时利用这些平均二级结构光谱的酰胺I、II和III带以及C -（α） - H酰胺弯曲振动来直接确定蛋白质二级结构。紫外拉曼方法似乎是对现有方法（如圆二色性、振动圆二色性和吸收光谱）的补充，在某些情况下还更优。此外，这些光谱不受困扰圆二色性、振动圆二色性和红外吸收测量的光散射伪影的影响。因此，将有可能检测胶束和其他散射介质中的蛋白质。
We have directly determined the amide band resonance Raman spectra of the "average" pure alpha-helix, beta-sheet, and unordered secondary structures by exciting within the amide pi-->pi* transitions at 206.5 nm. The Raman spectra are dominated by the amide bands of the peptide backbone. We have empirically determined the average pure alpha-helix, beta-sheet, and unordered resonance Raman spectra from the amide;resonance Raman spectra of 13 proteins with well-known X-ray crystal structures. We demonstrate that we can simultaneously utilize the amide I, II, and III bands and the C-(alpha)-H amide bending vibrations of these average secondary structure spectra to directly determine protein secondary structure. The UV Raman method appears to be complementary, and in some cases superior, to the existing methods, such as CD, VCD, and absorption spectroscopy. In addition, the spectra are immune to the light-scattering artifacts that plague CD, VCD, and IR absorption measurements. Thus, it will be possible to examine proteins in micelles and other scattering media.