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Integrated Structural Biology Grenoble

Contact person(s) related to this article / FAVIER Adrien

Fast characterization of protein samples using liquid-state NMR

This platform can be used to characterize:

The degree of folding of a protein
This is achieved by looking at the presence of amide and methyl proton resonances outside the random coil chemical shift regions in 1H 1D-NMR spectra or in 1H-15N correlated 2D spectra if the protein is 15N labeled.

The quantitative detection of the structural compactness of the protein.
This is achieved by recording 1D or 2D HET-SOFAST experiments if the protein is 15N labeled. These experiments allows to measure a parameter (λNOE ) which is related to the compactness of the protein. A low value of λNOE corresponds to a well-folded protein whereas a higher value reveals flexible regions. A single λNOE value is measured for the whole protein in the 1D HET-SOFAST experiment. With the 2D version, a λNOE parameter is measured for each residue in the protein

The oligomerisation state of the protein
Through the measure of its rotational diffusion (τc ) if the protein is 15N labeled.

The interaction of the protein (or acid nucleic) with its partner in solution.

Report example

Key words

Protein Folding, interaction, 1D NMR, 2D NMR, SOFAST NMR, HET-SOFAST NMR

Contact

Adrien FAVIER

Dedicated staff

- Adrien FAVIER

Specific equipment

600 MHz spectrometer equipped with HCN cryogenic probe

Quality control of the spectrometer’s specifications is done quaterly and a standard sample is used in case of dysfunction.

Website

http://www.ibs.fr/content/ibs/presentation/platform/quality

Access mode

PSB users

Cost

Free of charge for the PSB users

Location

IBS LRMN

How to make a request ?

Use this online request form

Additional information

Delay 1 week.
The user comes to the NMR lab with its sample and stays with the local operator during the experiments and the analysis (about 1 to 3 hours)

Sample requirement:
Toxic and/or radioactive samples are not accepted. Only the biological samples whose biohazard level is 1 (non-pathogenic) will be accepted.

The minimum concentration should be about 100 µM. The sample volume is 600 μl. The NMR tube is furnished by the NMR lab. The most suitable buffers for NMR are phosphate and deuterated ones but the following buffers can also be used:
- Na Acetate, pH=4.5-5.5
- Na Acetate, pH=5.5
- Na Citrate, pH=4.7-5.5
- NH4 Acetate, pH=7.3
- MES, pH=5.8-6.5
- TRIS, pH=7.5-8.5
- HEPES, pH=7.0-8.0
- Bicine, pH=8.5-9.0
- Cacodylic acid, pH=6.5

The salt concentration should be kept below 400 mM if possible. Traces of iron or paramagnetic buffer should be absent.

Aknowledgments

Please, add the following aknowledgments on your posters and publications :

« We thank Eric Condamine and/or Adrien Favier, from the IBS platform of the Partnership for Structural Biology and the Institut de Biologie Structurale in Grenoble (PSB/IBS), for the NMR access and the support provided »