Protein Secondary Structure Determination Using Drop Coat Deposition Confocal Raman Spectroscopy

The accurate determination of protein structure is integral to the medical and pharmaceutical communities' ability to understand disease and develop drugs. Current techniques (such as circular dichroism [CD], infrared [IR] spectroscopy, and Raman spectroscopy) for protein structure prediction provide results that can be poorly resolved, while high-resolution techniques (such as nuclear magnetic resonance [NMR] and X-ray crystallography) can be both costly and time-consuming. This work proposes the use of drop coat deposition confocal Raman (DCDCR) spectroscopy coupled with peak fitting of the amide I spectral region (1620-1720 cm(-1)) for the accurate determination of protein secondary structure. Studies conducted on bovine serum albumin (BSA) and ovalbumin show that the predictions of secondary structure content within 1% of representative crystal structure data is possible for model proteins. The results clearly demonstrate that DCDCR has the potential to be effectively used to obtain accurate secondary structure distributions for proteins.