Structural Differences of Proteins Between Solution State and Solid State Probed by Attenuated Total Reflection Fourier Transform Infrared Spectroscopy
Structural Differences of Proteins Between Solution State and Solid State Probed by Attenuated Total Reflection Fourier Transform Infrared Spectroscopy
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DOI：
10.1366/000370209787944316
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发表时间：
2009-04-01
影响因子：
3.5
通讯作者：
Cai, Shuowei
中科院分区：
文献类型：
--
作者：
Sachdeva, Amita;Cai, Shuowei
建立了一种结合衰减全反射（ATR）采样技术的傅里叶变换红外（FIAR）光谱分析方法，用于分析固态和溶液态蛋白质的二级结构。该方法已被应用于分析蛋白质在溶液状态和固体状态下的结构差异。对于α-螺旋占优势的蛋白质，β-折叠结构在固态下显著增加，α-螺旋结构显著减少。对于β折叠占优势的蛋白质，β折叠结构在固态下仅适度增加。当蛋白质重新溶解在溶液中时，它们的结构被重新自然化为它们的天然结构，正如它们在溶液状态下的结构与在溶液状态下通过X射线晶体学或其他光谱方法确定的结构相似的事实所表明的那样。ATR采样技术避免了FAAR光谱中固体样品的传统溴化钾（KBr）圆盘法所需的高压和化学品。我们的方法从这项研究中，证明ATR采样更适合于分析蛋白质结构的固体状态。
A Fourier transform infrared (FIAR) spectroscopic method combined with an attenuated total reflection (ATR) sampling technique has been developed to analyze protein secondary structure in both solid and solution states. The method has been applied to analyze the protein structural differences between solution state and solid state. For alpha-helix dominant proteins, beta-sheet structures increase significantly in the solid state, with significant decrease in alpha-helical structures. For beta-sheet dominant proteins, beta-sheet structures increase only moderately ill the solid state. When proteins are re-dissolved in solution, their structures are re-natured to their native structures, as suggested by the fact that their structures in solution state are similar to those determined by X-ray crystallography or other spectroscopic methods in solution state. The ATR sampling technique avoids the high pressure and chemicals that are needed for the conventional potassium bromide (KBr) disc method for solid samples in FAAR spectroscopy. Our approach from this study, demonstrated that ATR sampling is more appropriate for analysis of protein structures in the solid state.