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.
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
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 :
« This work used the platforms of the Grenoble Instruct-ERIC center (ISBG; UAR 3518 CNRS-CEA-UGA-EMBL) within the Grenoble Partnership for Structural Biology (PSB), supported by FRISBI (ANR-10-INBS-0005-02) and GRAL, financed within the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003) »
Terms of use
Terms of the NMR platform
- The experiments will be conducted by a local operator
- The samples are not stored
- The user can ask the local operator the raw data of the experiment and a report in pdf format will be delivered for standard experiments.
- The raw data are conserved for 2 years.
- The user agrees to acknowledge the platform when publishing and to communicate the reference to the NMR platform contact.
- The “Material & Method” section will be provided without co-authoring condition for standard experiments.