UE Structure determination of biological macromolecules

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:

  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                                                  

    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                                                  

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.