Near infrared spectroscopic characterisation of secondary structure content of proteins

摘要

Near infrared (NIR) spectroscopy was investigated as a non-destructive and rapid method to characterise the secondary structures of proteins in solid state. The absorption spectra of 11 reference proteins (0-46% α-helix, 6-73% β-sheet) were obtained using an Antaris NIR Analyser and quantitatively analysed using a chemometric software program to correlate the NIR spectral data to the secondary structure content obtained from X-ray crystallography and Raman spectroscopy. Simple linear regression analyses of the normalised second derivative NIR spectra indicated that 6289cm{sup}(-1) (R{sup}2 = 0.99, RMSEC=2.6) and 4602cm{sup}(-1) (R{sup}2=0.96, RMSEC=5.6) were highly sensitive wave number regions for β-sheet structure and the calibration models were highly predictive for three additional proteins which served as external validation standards (average error 2% and 3%, respectively). The second derivative NIR spectra at 4602cm{sup}(-1) was also found to be sensitive to α-helical content (R{sup}2 = 0.93, RMSEC=5.8) and predictive of the validation standards (average error 6%). The results demonstrate the potential of NIR spectroscopy as a rapid and non-invasive tool for quantification of secondary structure content of proteins in solid state.