CERM/CIRMMP provides state of the art instrumentation and expertise to perform the most comprehensive array of experiments needed for the structure and dynamic characterisation of biological macromolecules and their complexes. All the standard pulse sequences for spectroscopic, structural and dynamical characterisation are available for attaining fundamental atomic level information. CERM/CIRMMP have developed 13C direct detection protocols for “protonless” NMR experiments and for in-cell NMR spectroscopy, and tailored pulse sequences for structural determination of paramagnetic systems.
The infrastructure is equipped with high and ultra-high field spectrometers equipped with cryoprobes (1.2 GHz TCI cryo, 950 MHz TCI cryo, 900 MHz TCI cryo, 700 MHz TCI cryo, 700 MHz TXO cryo, 500 MHz TCI cryo), heteronuclear detection probes at 600 MHz for low frequency nuclei to meet the most disparate experimental needs, liquid handler system and autosamplers for metabonomics and ligand screening at 400, 600 and 700 MHz. Special accessories such as high power probes (short hard pulse) at 400 and 600 MHz for paramagnetic NMR. All NMR instruments are state-of-the-art digital spectrometers of the Bruker Avance NEO/III series.
To get structural information on proteins by solution NMR, isotopically labelled samples, usually 15N and/or 15N/13C, are required. For proteins with MW > 20 kDa, deuteration is also needed. In general, the degree of deuteration required is largely dependent on spectral quality, the size and modularity of the system studied and the types of experiment performed. However, perdeuteration (>90 %) are usually required for proteins with molecular weights of the order of 35 kDa or greater. A screening with several buffers (with different pHs, ionic strength and containing chemicals such as detergents, protease inhibitor cocktails, arginine etc.) is recommended to optimise NMR samples.
The support might include sample preparation, instruments setup, data acquisition and data analysis. Based on the visiting scientist expertise assistance and supervision will be assigned.
For remote access, if the samples are not shipped in the NMR tube, our staff takes care to transfer it from the vial to the tube; air-sensitive samples will be handled in an oxygen-free chamber. Our staff will operate following the protocol for operations (air/light sensitivity, temperature of storage, etc…) provided together with the samples.
The visiting scientist must communicate the scientific plan of the experiments and the contacts to arrange the visit. On average a visit is one week of instrument time and it can be arranged as a physical visit to the facility or by mailing-in the samples. Some projects might require more time, up to several weeks, depending on the complexity of the work to be performed and the experience of the visiting scientist in the field.
Contact the facility staff before submitting your proposal, so that the access request is in agreement with the scientific plan of the experiments.
