Ateneo - UPO

The course is divided into two modules.
The first part of the first module provides an introductory leveling section where basic principles and notions of proteins and enzymes, their properties and functions will be discussed.
The second part will descibe the main methods for the study and analysis of proteins with an in-depth analysis of the techniques for structural investigation (mainly X-ray diffraction and notions of electron cryomicroscopy).
In the third part, basic principles of drug design and protein engineering on a rational basis will be presented and discussed. Moreover, scientific publications relevant and more representative of the application aspects of protein science will be presented and discussed.

In the second module, the basic principles of protein engineering and the related reference techniques will be addressed. The physical-chemical methods for the study of protein-protein and protein-ligand molecular interactions will also be discussed.

DonaldVoet, Judith G Voet, Charlotte W Pratt. PRINCIPIDI BIOCHIMICA, 2017. Zanichelli.
Introduzione allo studio delle proteine. Duranti; Zanichelli
Donald Voet, Judith G Voet, Charlotte W Pratt. FONDAMENTI DI BIOCHIMICA. IVedizione, 2017. Zanichelli
David L Nelson, Michael M Cox. PRINCIPI DI BIOCHIMICA DI LEHNINGER. VII edizione, 2018. Zanichelli    
Mike Williamson. "Come funzionano le proteine" Edizione Italiana a cura di Martino Bolognesi, 2020. Zanichelli.

The course provides the student with an in-depth study of the fundamentals of protein biochemistry acquired during the first level degree, with particular attention to the structure-function relationships of biological macromolecules, structural biology and biophysical methods for the characterization of proteins.

Basic knowledge of general chemistry, organic chemistry, physics and biochemistry.

Lesson ex-cathedra with the slides presentation, and movies. Execution of problems and quizzes in the classroom, discussion on literature data. Students can also use the recommended textbooks to deepen the topics discussed in the classroom

In itinere test will be carried out during the course.

See the description of the general course.

Basic principles of protein science:
-Amino acids and proteins: chemical and structural properties
-Peptide bond and protein structure
-Proteins as information molecules: sequence and database analysis

Structural analysis techniques:
Structural biology: spectroscopic and diffrattometric techniques for structural protein investigation
Protein visualization programs
Structural protein prediction
Molecular modelling

"Case studies" and application implications of Protein Science:
-Protein science and drug design: principles, examples and discussion of scientific publications
-Protein Sciences and Protein Engineering: Principles, Examples and discussion of scientific publications

PROTEIN ENGINEERING

- Introduction to the production of recombinant proteins
- Cloning techniques and expression vectors
- Site-directed and random mutagenesis
- Chimeric proteins
- Rational protein design: theoretical principles and experimental methods
- Selection and evaluation of protein variants: biochemical assays and desired property analysis.

CHEMICAL-PHYSICAL METHODS FOR PROTEIN ANALYSIS:
-Chromatographic techniques for protein purification
- Salting in/out.
- Methods for assessing the concentration and purity of proteins.
Methods for assessing protein stability and folding:
- Differential scanning fluormetry and differential scanning calorimetry
- Circular dicroism
- "Light-Scattering" for the study of the quaternary structure of proteins

-Methods for the analysis of protein-protein and protein-ligand interactions:
- Isothermal titration calorimetry
- Surface plasmon resonance
- Microscale thermophoresis

The student should be able to argue and discuss the basic principles of the reactivity of biological macromolecules and the structure-function relationship of proteins. The student should also know the reference techniques for the biochemical and structural characterization of biological macromolecules.