UE Structure determination of biological macromolecules

Diplômes intégrant cet élément pédagogique :

Descriptif

 

The course will present practical and theoretical aspects of the classical approaches used to determine the structure of macromolecules by X-ray crystallography and Nuclear Magnetic Resonance spectroscopy. The program of this course is described below:

 

q  Part I - Crystallography - 20h

  • Crystallization techniques                                                                            
  • Crystal symmetry and space groups                                                                      
  • Diffraction                                                                                                    
  • Structure factors, reciprocal space etc                                                      
  • Tutorial: Data treatment                                                                                          
  • Phasing – MIR, SAD, MAD                                                                        
  • Molecular replacement, crystallographic symmetry                                              
  • Tutorial: MAD Phasing                                                                                              
  • Tutorial: Molecular replacement                                                                             
  • Refinement                                                                                                  
  • Tutorial: Model building and refinement                                                                  
  • Practical lab: Crystallization on a PSB platform                                                                 
  • Practical lab: X-ray data collection on a ESRF beamline                                                  

 

q    Part II – Nuclear Magnetic Resonance-20h

  • NMR principles: active nuclei, magnetic field, radiofrequency excitation,
               return to equilibrium                                                                                     
  • NMR observables in the spectra of biomolecules (chem. shift, scalar
               couplings,  linewidth)                                                                        
  • NMR observables: measurement                                                               
  • Practical lab on IBS-NMR platform: data collection
  • The steps to structure determination: sample preparation, isotopic labeling
  • The steps to structure determination: assignment   
  • The steps to structure determination: extraction of structural parameters 
  • Tutorial: Data analysis                                                                                 
  • Structure calculation: principles                                                                              
  • Practical lab: Protein structure calculation                                                  

 

Compétences visées

Decision making in structure determination of biomolecules, expertise in experimental structure determination by X-ray crystallography and liquid-state NMR spectroscopy, critical analysis of structural models at atomic resolution.

Informations complémentaires

Langue(s) : Anglais