Nuclear Magnetic Resonancehttps://www.renafobis.fr/ecoles-thematiques/ecole-doleron/ecole-oleron-2015/rmnhttps://www.renafobis.fr/@@site-logo/paris_rainbow-renafobis.png
After a rapid introduction on NMR spectroscopy principles, chemical shifts will be analyzed as a source of secondary structure element and a tool to decipher the localization of interacting partners, together with binding constant parameters. Relaxation will be introduced as a tool to analyze ps-to-ns motions, as well as ms--to-us dynamics. Scalar couplings and dipolar interactions will finally be described as a source to extract orientational restraints and dynamical information.
In a context of deciphering, understanding and predicting the function of biological macromolecules, NMR probes the electronic environment of nuclei and their time evolution over a uniquely wide range of time-scale and at atomic resolution. The relevant NMR parameters and motional time scales will be described through the study of biological processes such as protein folding, functional disorder, catalytic, regulatory or signalization processes, or molecular recognition.
Practical Work
The practical work aims at the qualitative and quantitative analysis of 15N-HSQC spectra of several proteins. This will illustrate how this kind of spectra allows to rapidly obtain structural and dynamic informations on proteins under various conditions and on its interactions with partners.
Contact: Ewen Lescop (ewen.lescop_at_cnrs.fr) Softwares to be installed :
(1) CCPNMR Analysis 2.4.2 (http://www.ccpn.ac.uk/downloads/stable) Download the version corresponding to your OS here: "Analysis pre-compiledbinaries (includes all the required libraries)" and follow the instructions for installation.
(2) PyMOL. Follow the instructions for the installation in the softwares for XRay.
For MacOSX users: CCPNMR requires X11. For that, it is recommended to install XQuartz
Self-assembly of multiple copies of protein subunits into large supramolecular complexes plays a key role in many cellular processes, notably in bacterial virulence, neurodegenerative disease-related mechanisms, and in biomolecular machines. 3D structures of these macromolecular assemblies are still rare as they are usually non-cristalline and insoluble and therefore recalcitrant to X-ray diffraction and solution NMR techniques. I will present recent developments in Solid-State NMR (SSNMR) to experimentally determine supramolecular interactions at the subunit interfaces, ultimately aiming at atomic resolution structures of biological macromolecular assemblies. I will discuss the combination of SSNMR methods with density maps obtained by cryo-electron microscopy and advanced modeling methods in an integrative multi-technique approach.
