Protein Dynamics By Raman Spectroscopy

[carxtal]

Can one monitor dynamics of a specific protein domain (helix, strand etc) by Raman Sp? If one labels a specific residue with an isotope, like in NMR, can one use that to monitor the dynamics of the region that encompasses this residue?

 

[wajagawa]

From my understanding (or lack thereof), you do not need any specific modifications to a protein in order to study a specific area in dynamic raman spec. studies (provided that the area(s) of interest are identifiable and show strong enough absorbance/signal). However, more often than not dynamics studies in proteins that monitor specific areas of a protein utilize some form of mutant proteins (some truncated, a residue here and/or there swapped, etc.).
Perhaps, this study of the folded and unfolded states of OmpA may be of use to you http://galileo.ucsd.edu/pdf/sanchez_jpcb_08.pdf

Please let me know if this helps.

 

[carxtal]

Thanks much!
Therefore, if i have 2 mutants to a WT protein I should be able to measure dynamics of these 3 proteins. Can I localize the analysis of the dynamics to a specific domain? Say a single helix? Can one perform a differential analysis to compare WT->mutant 1 and WT->mutant2? Simply put, can one subtract spectra in a statistically significant manner?
Fianlly, I understand that residues with aromatic sidechains like tryptophans are more significant spectra. Will it be an issue of the domains in question do not have such aromatic residues?
Thanks again!