Quantification of casein phosphorylation with conformational interpretation using Raman spectroscopy.
Raman spectroscopy is emerging as a powerful method for obtaining both quantitative and qualitative information from biological samples. One very interesting area of research, for which the technique has rarely been used, is the detection, quantification and structural analysis of post-translational modifications (PTMs) on proteins. Since Raman spectra can be used to address both of these questions simultaneously, we have developed near infrared Raman spectroscopy with appropriate chemometric approaches (partial least squares regression) to quantify low concentration (4 microM) mixtures of phosphorylated and dephosphorylated bovine alpha(s)-casein. In addition, we have used these data in conjunction with Raman optical activity (ROA) spectra and NMR to assess the structural changes that occur upon phosphorylation.
Jarvis
Roger M
RM
Manchester Interdisciplinary Biocentre, 131 Princess Street, Manchester, UKM1 7ND. roger.jarvis@manchester.ac.uk
Blanch
Ewan W
EW
Golovanov
Alexander P
AP
Screen
James
J
Goodacre
Royston
R
D000818:Animals; D002138:Calibration; D002364:Caseins / Q000737:chemistry / Q000378:metabolism*; D002417:Cattle; D016018:Least-Squares Analysis; D009682:Magnetic Resonance Spectroscopy; D010766:Phosphorylation; D011487:Protein Conformation; D019265:Spectroscopy, Near-Infrared; D013059:Spectrum Analysis, Raman / Q000379:methods
D002364:Caseins