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 such as chemical shift assignment, 15N relaxation measurements, structure elucidation and protein-ligand interactions. CERM/CIRMMP has 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 as well as for the study of Intrinsically Disordered Proteins and Regions (IDPs and IDRs).
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login to contact To get structural information on proteins by solution NMR, 15N and/or 15N/13C isotopically labelled samples are usually required. For very large folded proteins, deuteration might also be needed.
Crucial for optimal NMR sample are protein concentration and buffer composition. Depending on the structural and dynamic features of the sample and on the scientific question of the applicant, optimal protein concentration is usually in the range of hundreds of micromolar or higher.
Buffers with low salt concentration, when possible, are often preferred over high-salt-concentration buffers.
The applicant is always welcome to directly contact the scientific staff of CERM/CIRMMP prior access proposal submission for the evaluation of the feasibility of the project.
The support, when requested, includes sample handling, setup of NMR experiments, data acquisition, data analysis and data sharing. Based on the expertise of the visiting scientist, 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 them to the NMR tube (3mm, 5mm, shigemi, shaped tubes are available); air-sensitive samples will be handled in an oxygen-free chamber. Our staff will operate in concert with the applicant 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.
Contact the facility staff before submitting your proposal, so that the access request is in agreement with the scientific plan of the experiments.