How can we compare stability of two proteins with the help of NMR spectroscopy? what are the traditional NMR techniques that we use for this purpose?
The NMR spectroscopy is a powerful technique related to atomic level. Concerning protein denaturation, it's not sufficient alone but need other techniques.
Take a look at this mini review, it may help you in this subject.
Article Modern Technologies of Solution Nuclear Magnetic Resonance S...
NMR spectroscopy is a powerful technique for structural studies of chemical compounds and biomolecules such as DNA and proteins. Since the NMR signal sensitively reflects the chemical environment and the dynamics of a nuclear spin, NMR experiments provide a wealth of structural and dynamic information about the molecule of interest at atomic resolution. In general, structural biology studies using NMR spectroscopy still requires a reasonable understanding of the theory behind the technique and experience on how to recorded NMR data. Owing to the remarkable progress in the past decade, we can easily access suitable and popular analytical resources for NMR structure determination of proteins with high accuracy. Here, we describe the practical aspects, workflow and key points of modern NMR techniques used for solution structure determination of proteins. This review should aid NMR specialists aiming to develop new methods that accelerate the structure determination process, and open avenues for non-specialist and life scientists interested in using NMR spectroscopy to solve protein structures.
1. ·The use of osmolytes to facilitate protein NMR spectroscopy. 1993, Volume 3, Issue 5, pp 597–600
2. Sibille et al.Comparative NMR study on the impact of point mutations on protein stability of Pseudomonas mendocina lipase ;Protein Science (2006), 15:1915–1927.
Hi Sujeesh,
As far as I know, stability is usually defined by a melting temperature as obtained from DSC curves. Maybe it is best in NMR to simply go for a temperature ramp and look what happens. You can try to construct a Vant Hoff-type plot and then you will see e.g. a phase transition. Simply try some things ...
We also encountered a similar issue with EPR spectroscopy and were able to make some stability conclusions from ligand interactions with polymers.
Have a look at this one (doi:10.3390/polym9080324 ):
Article Probing the Nanoscopic Thermodynamic Fingerprint of Paramagn...
Give it a try ...
Good Luck !
Dear Sujeesh,
two short add-ons to all correct comments from above:
- Temperature dependent DOSY (no isotope labeling needed) might also be a option, because "molten/denatured" proteins have less dense packing. This was evaluted many years back by Wilkins et al
Article Hydrodynamic Radii of Native and Denatured Proteins Measured...
- Overall measure for presence of higher-order structure was also monitored at natural abundance by use of 1H-13C spectroscopy in nice work by Luke Arbogast recently
Article Mapping Monoclonal Antibody Structure by 2D 13 C NMR at Natu...
Cheers
Alfred
P.S I agree with Jörg, that DSC is probably the better and more established method to study thermal (in)stabiliy!
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