SECONDARY STRUCTURE AND DOSAGE OF SOLUBLE AND MEMBRANE-PROTEINS BY ATTENUATED TOTAL REFLECTION FOURIER-TRANSFORM INFRARED-SPECTROSCOPY ON HYDRATED FILMS
SECONDARY STRUCTURE AND DOSAGE OF SOLUBLE AND MEMBRANE-PROTEINS BY ATTENUATED TOTAL REFLECTION FOURIER-TRANSFORM INFRARED-SPECTROSCOPY ON HYDRATED FILMS
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DOI：
10.1111/j.1432-1033.1990.tb19354.x
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发表时间：
1990-10-24
期刊：
影响因子：
--
通讯作者：
RUYSSCHAERT, JM
中科院分区：
文献类型：
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作者：
GOORMAGHTIGH, E;CABIAUX, V;RUYSSCHAERT, JM
衰减全反射傅里叶变换红外光谱的可溶性和膜蛋白质的磷脂双层薄膜水合膜显示提供有用的信息，蛋白质的二级结构。通过傅立叶自去卷积，然后由曲线拟合的氘代样品的酰胺I带的分析是由一个新的程序，其中所有的输入参数是由计算机产生的，而不是由调查员。该分析的结果提供了α-β的正确估计。螺旋和β-当X射线结构作为参考时，具有8.6%的标准偏差的片状结构含量。我们还表明，通过傅立叶自解卷积/曲线拟合程序和磷脂酰基链的解决的不同的二级结构的取向可以同时评估在脂质双层中重构的膜蛋白。对于膜蛋白的重建特别感兴趣的是，可以从红外光谱准确快速地确定脂质蛋白比。
Attenuated total reflection Fourier-transform infrared spectroscopy of thin hydrated films of soluble and membrane protein included in a phospholipid bilayer is shown to provide useful information as to the secondary structure of the protein. The analysis of the amide I band of deutered samples by Fourier self-deconvolution followed by a curve fitting was performed by a new procedure in which all the input parameters are generated by the computer rather than by the investigator. The results of this analysis provide a correct estimation of the .alpha.-helix and .beta.-sheet structure content with a standard deviation of 8.6% when X-ray structures are taken as a reference. We also show that the orientation of the different secondary structures resolved by the Fourier self-deconvolution/curve-fitting procedure and of the phospholipid acyl chains can be simultaneously evaluated for membrane proteins reconstituted in a lipid bilayer. Of special interest for reconstitution of membrane proteins, the lipid protein ratio can be accurately and quickly determined from the infrared spectrum.