Evolution of quantitative methods in protein secondary structure determination via deep-ultraviolet resonance Raman spectroscopy

Abstract

Deep-ultraviolet resonance Raman (DUVRR) spectra is sensitive to secondary structural motifs but, similar to circular dichroism (CD) and infrared spectroscopy, requires the application of multivariate and advanced statistical analysis methods to resolve the pure secondary structure Raman spectra (PSSRS) for determination of secondary structure composition. Secondary structure motifs are selectively enhanced by different excitation wavelengths, a characteristic that inspired the first methods for quantifying secondary structures by DUVRR. This review traces the evolution of multivariate methods and their application to secondary structure composition analyses of proteins by DUVRR spectroscopy from the first experiments using two-wavelengths, and culminating with recent studies utilizing time-resolved DUVRR measurements.

Improvement of the in vivo cellular repopulation of decellularized cardiovascular tissues by a detergent-free, non-proteolytic, actin-disassembling regimen

“…This is all the more important in the context of biomaterials undergoing decellularization, as ideal decellularization regimens would remove all cellular proteins while preserving all extracellular proteins including their conformation. The increased Raman amide I band at 1659 cm‐1 in BIO‐decellularized pAV leaflets may indicate a higher number of proteins in alpha‐helix or beta‐turn conformation as compared with DET grafts, whereas exact interpretation of the secondary protein structure is still a matter of debate in the literature (Pelton and McLean, ; Roach et al , ; Spiro and Gaber, ). However, the findings point at ordered molecule structures, especially in BIO prostheses, indicating preserved protein conformation.…”

Self-assembled peptide nanostructures for functional materials

“…However, conventional Raman spectroscopy lacks the spatial resolution fordetailed characterizations of the peptide nanostructures duelow signal to noise ratios at dilute conditions,and different approaches have been developed to improve the scattering intensities for the analysis. UV resonance Raman spectroscopy, which enhances the frequencies in vibrational amide modes including amide III and C α -H bands were applied to selectively monitor the secondary, tertiary or quaternary organization of the peptides in solution [255][256][257][258]. The relatively low Raman signal of the peptide assemblies wasalso increased via surface-enhanced Raman scattering (SERS) using the localized surface plasmon resonances of the metallic clusters in liquid or solid environments [259,260].…”

Determining the Secondary Structure of Membrane Proteins and Peptides Via Electron Spin Echo Envelope Modulation (ESEEM) Spectroscopy

“…However, it requires milligram scales of isotope-labeled protein or peptide samples and days to weeks of data-acquisition time while still suffering from low sensitivity. Other methods such as FT-Raman spectroscopy, ATR FT-IR, and continuous-wave EPR dipolar wave analysis also can provide secondary structure information (Carbonaro & Nucara, 2010; Roach, Simpson, & JiJi, 2012). Data obtained by these methods are sometimes ambiguous and often require extensive data analysis.…”