Am in Peptide / Protein Research, may i know what useful analytical information we are getting from 13C & 1H NMR spectroscopy analysis?
Somebody getting NMR for smaller proteins, any useful information we will get it from this?
In general, NMR is very informative and powerful technique. You can estimate the relative amount of your peptide/protein, the purity of the sample, or even the fold of the peptide/protein. You can also do interaction studies or stability tets.. Recording simple 1H spectrum can already tell you such information. But there is much more NMR can do. I would recommend to start with this very nice book:
NMR of Biomolecules: Towards Mechanistic Systems Biology
Editor(s):
- Prof. Dr. Ivano Bertini
- Kathleen S. McGreevy
- Prof. Giacomo Parigi
In general, NMR is very informative and powerful technique. You can estimate the relative amount of your peptide/protein, the purity of the sample, or even the fold of the peptide/protein. You can also do interaction studies or stability tets.. Recording simple 1H spectrum can already tell you such information. But there is much more NMR can do. I would recommend to start with this very nice book:
NMR of Biomolecules: Towards Mechanistic Systems Biology
Editor(s):
- Prof. Dr. Ivano Bertini
- Kathleen S. McGreevy
- Prof. Giacomo Parigi
Mr. Sebastian,
The quantitative and 3D structural information gained by NMR is useful, but not exact; even it is not accurate, talking about 3D molecular conformations of the peptides, due to the fact that the accuracy of the stuctural parameters such as dihedral angles is at about 4o. The latter value represents a significant deviation, since there are a set of 3D molecular conformations and electronic structures of an analyte with very close free Gibbs energy values which can be significantly different; however, within the framework of these 4o deviation of the corresponding geometry parameter.
Currently, exact 3D structural analysis of molecular and electronic structures of the analytes can be carried out only by mass spectrometry; however, explicitly, within the framework of our own-authored (to me and my co-author's one) innovative stochastic dynamic concept and model formulas.
In this context, please, consider the following review-article [1]. It discusses briefly the capability of NMR and CD spectroscopy of gaining 3D structural information of analytes compared with the capability of the methods of mass spectrometry within the framework of our theory and model equations.
[1] Reviews in Analytical Chemistry, Band 38: Heft 2 (2019)
A stochastic dynamic mass spectrometric diffusion method and its application to 3D structural analysis of the analytes; Bojidarka Ivanova, Michael Spiteller
For oligopeptides 1H and 13C NMR is a useful technique. You can estimate the purity of the sample, its stability in solution, or follow the kinetics of reactions with the solvent.
There are several reviews of Ivano Bertini on the subject for instance “NMR of biomolecules”, but also too many recent articles analysing NMR spectra
Dear Arun Manakkal Sebastian
Peptide purity mainly depends on HPLC Then Mass spectrum
But via NMR Spectrum If small peptide means some structural information like organic compound may confirm out of that the side-chain group environment may identify.
For long sequences don't know what we can find
Thnaks
Dr Athira Nair
Dear Arun Manakkal Sebastian
NMR is very used full technics for secondary structure determination in solution state in the peptide. if you don't have crystal structure NMR can help for the same.
Thanks
Bibhu
One of the best NMR pulse sequence to study peptides is NOESY 2D. You can find many articles with sugestions to choose the best way to setup de aquisition parameters and, very important, how to prepare the simples. My second sugestion: before nmr make a determination of aminoacids sequence. Best regards
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