Méthodes de la biologie structurale

Description

Understand how biomolecular structures are determined by X-ray diffraction and NMR spectroscopy. What are the observables and limiting steps? Be able to apply NMR experiments to biomolecules for assignment and structure analysis. Judge the quality of a 3D structure. Comparison of technologies including X-ray, NMR and cryo-electron microscopy.Importance of structural approaches in chemical biology and biology.

Getting structure to learn biology. Fundamentals of macromolecular crystallography: crystal geometry, diffraction basics, getting crystals, data collection and processing. Phasing and interpretation of an electron density map. Refinement, structure validation and analysis.
Spin manipulation by simple radiofrequency pulse experiments and spectra analysis after Fourier transformation. Analysis of 2D NMR spectra of biomolecules such as COSY, TOCSY, NOESY and heteronuclear experiments. Spectral assignment of proteins. How is structural information extracted from NMR spectra? Structure calculation from NMR data. Isotopic labelling for NMR.
 

Compétences visées

• Understanding of concepts and methods presented in the class.
• Connect and integrate from different domains, in particular for 3D structures (X-ray, NMR, cryo-electron microscopy and other methods). What are the prerequisites to apply each technique, their specific properties and complementarity?
• Ability to analyse a PDB structural file (evaluation of a structure and quality control by the user).
• Analyse a biomolecular structure in its biological context.
• Analyse and interpret NMR spectra to establish a 3D structure.
• Understand NMR methods sufficiently to establish simple NMR pulse sequences.
• Understand the steps in structure determination. In an automated process know the decision points and the limiting steps.
• Organize and plan your work.
• Work in an independent manner, establish a time schedule and auto-evaluation.
• Ability of analytical and scientific thinking.
• Work as a team.
• Develop and formulate a rigorous causal reasoning.
• Oral presentation in public and the proper use of supports.
• Be able to think in an abstract manner.

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