Macromolecular structural biology

The course is focused on the 3D structure determination and analysis of proteins by using biocrystallographic and bioinformatic techniques. Experimental techniques for protein crystal growth will be described in details, with application to soluble and membrane proteins. X-ray diffraction on protein crystals and on solution samples (SAXS) will be treated extensively (X-ray sources, synchrotron light, atomic scattering factors, structure factors, the "phase problem", phasing by molecular replacement and heavy atoms, crystallographic refinement). Protein model validation techniques will be analyzed, together with the main features of the Protein Data Bank. Examples of structural proteomic projects will be provided, with emphasis on high-throughput applications of the biocrystallographic and bioinformatic methods learned during the course. In addition, the theoretical basis and the practical applications of dynamic light scattering and molecular dynamics techniques will be treated. The last part of the course will have a more practical approach. Students will solve by molecular replacement, refine, and analyze the 3D structure of a protein. During this practical experience they will learn how to apply techniques and software for protein sequence alignment, secondary structure prediction, molecular modelling, crystallographic refinement and validation