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: Prof. 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: Prof. Luisa Iommarini)

Examples of applications of structural data for the study of protein function, including catalytic mechanisms, in some fundamental cellular processes:

• Control of cell cycle, mechanisms of regulation of proteins involved in cell cycle.
• Intracellular signalling (G-protein coupled receptors, RTK, nuclear receptors.
  
• Proteins involved in ATP production (structural arrangement of OXPHOS complexes and super-complexes, and thier catalytic mechanisms.
  

 

Module 3  (2 ECTS, 30 hrs, practical lab, teacher: Prof. Luisa Iommarini)

• Sample preparation for denaturing polyacrylamide gel electrophoresis SDS-PAGE: lysis of bovine heart mitochondria and protein 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 bovine heart mitochondria and protein dosage. SDS-PAGE results analysis (6 hrs).
• Native solubilisation of respiratory chain super-complexes from bovine heart mitochondria and protein dosage (4 hrs).
• Blue-Native polyacrylamide gel electrophoresis (BN-PAGE) and in gel activity (IGA) of complexes/super-complexes (8 hrs). 
• Analysis of experimental results.

 

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 (some lectures will be given just on-line). 

 

 

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. In case of persistence of the COVID-19 emergency, exams will be oral on the Teams or Zoom platforms.

Teaching tools

Videoprojector, PC, internet. Lecture slides will be available.

Office hours

See the website of Maria Paola Turina

See the website of Luisa Iommarini

See the website of Luisa Iommarini