Studying the glycan moiety of RNase B by means of Raman and Raman optical activity.
Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules.
Department of Chemistry, University of Antwerp, Groenenborgerlaan 171, 2020 Antwerp (Belgium).
Carl
C
Mensch
Robert
R
Pendrill
Göran
G
Widmalm
Christian
C
Johannessen
D004187:Disaccharides / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry*; D020033:Protein Isoforms / Q000737:chemistry; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D012260:Ribonucleases / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D013059:Spectrum Analysis, Raman / Q000379:methods
D004187:Disaccharides; D011134:Polysaccharides; D020033:Protein Isoforms; D012996:Solutions; D012260:Ribonucleases; C029600:ribonuclease B; D012259:Ribonuclease, Pancreatic
Raman optical activity; Raman spectroscopy; carbohydrates; glycoproteins; ribonuclease