85304 - Structural Biology

Learning outcomes

By the end of the course the student is provided with basic knowledge pertaining X-ray crystallography, NMR and cryoelectron microscopy applied to the structural characterization of proteins and nucleic acids. By the end of the hands-on lab the student is able to use programs for the interactive visualization and analysis of molecular structures and related data.

Course contents

Module 1  (3 ECTS, 24 hrs, teacher: M. Paola Turina)

• Elements of protein structure and protein folding.
  
• Overview of the experimental methods applied to obtain the structure at atomic resolution of proteins and nucleic acids: X-ray crystallography, NMR spectroscopy, Cryoelectron Microscopy.
  
• Use of Molecular Visualization software.

 

Module 2  (3 ECTS, 24 hrs, teacher: Luisa Iommarini)

Examples of structure-function relationship in some fundamental cellular processes:

• Energy transduction, OXPHOS system, structural organization of OXPHOS complexes and supercomplexes, regulation of OXPHOS complexes.
• Intracellular signalling, G-protein coupled receptors, RTK, nuclear receptors, downstream effector kinases, regulation of their activity through phosphorylation and other post-translational modification, activation of DNA transcription upon specific stimuli
• Protein turnover, ubiquitination system and other proteolytic effectors.
• Control of cell cycle, mechanisms of regulation of proteins involved in cell cycle.

 

Module 3  (2 ECTS, 30 hrs, practical lab, teacher: Claudia Zanna)

• Sample preparation for denaturing polyacrylamide gel electrophoresis SDS-PAGE: lysis of mitochondria of bovine heart and proteins dosage (4 hrs).
• Mitochondria of bovine heart lysates separation by SDS-PAGE and OXPHOS subunits visualisation by western blotting (8 hrs).
• Native solubilisation of oxidative phosphorylation complexes from mitochondria of bovine heart and protein dosage. SDS-PAGE results analysis (6 hrs).
• Native solubilisation of respiratory chain super-complexes from mitochondria of bovine heart and protein dosage (4 hrs).
• Blue-Native polyacrylamide gel electrophoresis (BN-PAGE) and complexes/super-complexes in gel activity (IGA) (8 hrs).                   
  

Readings/Bibliography

Selected reviews, scientific articles and book chapters will be provided

Teaching methods

Modules 1 and 2: Teacher lectures and student involvement in PC-based activities.

Module 3: Hands-on lab activities, assisted by introductory lectures and tutorials.

 

Assessment methods

Written test and oral interview. The written test will consist of a few open-ended questions and a few multiple-choice questions. The oral interview will discuss the written test.

Teaching tools

Videoprojector, PC, internet, board.

Office hours

See the website of Maria Paola Turina

See the website of Luisa Iommarini

See the website of Claudia Zanna