043CM - BIOCRYSTALLOGRAPHY AND ELECTRON MICROSCOPY 2024
Schema della sezione
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Introduction to structural biology: the importance of structure-property relationships. Protein databases. Software for visualizing protein structures.
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Hypothetical mechanism of the enzyme, based on the crystallographic structures of the protein obtained in different states.
The video was prepared with UCSF Chimera starting from coordinate files available on PDB.
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This video was generated following the procedure described in the last slides of the introduction.
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Basics of protein structure: primary, secondary, tertiary, and quaternary structure, with examples. Geometric constraints and degrees of freedom. Post-translational modifications and co-factors. Motifs and domains. Membrane proteins.
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Expression of recombinant proteins for structural studies. Design of the protein construct. Selection of a suitable expression system. Main molecular biology techniques used in structural biology.
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Bioinformatics resource operated by the SIB Swiss Institute of Bioinformatics. It includes various bioinformatic tools, among which databases and online software for structural biology, medicinal chemistry, genomics, proteomics, and systems biology.
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This protein sequence database includes annotations about localization, function, structure, post-translational modifications, etc. The website also contains links to bioinformatics tools for sequence alignment (Align) and primary sequence homology searches (Blast).
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Purification techniques. Methods to increase the stability, conformational homogeneity, and crystallizability of a protein. Evaluation of the quality and purity of the sample. Analysis of the conformational stability. Analysis of the oligomeric state of the protein.
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Crystallization of soluble and membrane proteins. Thermodynamic and kinetic perspectives on protein crystallization. Main crystallization techniques and optimization strategies, including co-crystallization and soaking.
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Protein crystals. Symmetry point elements. Crystal symmetry. Crystal families and systems. Bravais lattices. Screw axes: rotation and translation. Fractional coordinates. Space groups. International Crystallographic Tables. Non-crystallographic symmetry. Miller indices and reciprocal lattice.
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X-ray diffraction: basic concepts about the physics of diffraction. Bragg's law. Ewald sphere and reciprocal lattice. Symmetry of the reciprocal lattice. Laue classes. Systematic absences. Anomalous scattering.
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Small home-made guide for beginners.