Determination of the secondary structure of proteins from the Raman amide I band: the reference intensity profiles method

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.

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