Medicine and Surgery

Teaching objectives: • know the structure and function of amino acids, oligopeptides and water-soluble vitamins • know the structure and function of fibrous proteins (collagen), myoglobin, hemoglobin • know the properties and function of enzymes and the control and regulation mechanisms of enzymatic activity • know some methodologies for studying and characterizing proteins • know the structure and function of carbohydrates and the bonds that are involved in polymeric structures and for the formation of complexes with proteins and lipids (glycoproteins and glycolipids) • know the structure and function of lipids. Role of lipids in the composition of biological membranes, classification of membrane lipids. Structure of membrane proteins and the bonds involved. Mechanism of transport across membranes • know how the structure of DNA and the bonds that stabilize it and that intervene in the specific interactions between DNA and proteins (structural and regulatory) are the basis of the control of gene expression. Preliminary notions: chemical composition of living matter. Properties of water. Biological buffer systems. Proteins: amino acids. Stereochemistry, classification, acid-base properties, isoelectric point. Peptide bond, structure and properties. Ramachandran chart. Glutathione. General structure and maturation of insulin. Structure of proteins: levels of organization. Domains and structural motifs. Collagen and its maturation. Vitamin C: structure and role. Pathologies associated with alterations in collagen. Alpha-keratins. Hemoglobin and myoglobin, structure and function. Saturation curve. Hill equation. Cooperativeness and its structural bases. Bohr and Haldane effects. Allosteric effectors (2,3-BPG, CO2, H+, Cl-). Transport of CO2 in the blood. Allosteric models. Hemoglobinopathies. Introduction to proteomics. Protein folding. Chemical and physical denaturants. Proteins that assist folding. Chaperones and chaperonins. The hydrophobic effect. Anfinsen's experiment and Levinthal's paradox. Biochemical bases of neurodegenerative diseases caused by misfolding. Principles of biochemical methodology. Methods of purification and characterization of macromolecules: chromatography, electrophoresis, spectrophotometry. Isoelectrofocusing and SDS-PAGE (1D and 2D). Notes on mass spectrometry and crystallography, Enzymes. Enzyme nomenclature and classification Catalysis and free energy of activation. General properties of enzymes: reaction speed, specificity and stereospecificity of the reaction, regulation. Chemical kinetics: reaction rate, reaction order, first order and second order specific rate constants. Enzyme kinetics: The pre-steady state (outline) and the steady state. Michaelis-Menten equation. Meaning of KM and Vmax (kcat) and specificity constant. Graphical determination of steady state parameters. Linear transformations: Lineweaver-Burk. Enzyme inhibition: competitive, mixed incompetitive, inhibition. Catalytic mechanisms: acid-base, covalent and metal-mediated (carbonic anhydrase, lysozyme, serine protease and aspartyl protease). Water-soluble vitamins. Group B vitamins. Sources of vitamins and pathologies associated with vitamin deficiencies. Vitamin coenzymes and their structure. Blood clotting – Platelet aggregation and release of effectors. Factors involved in coagulation: serine proteases, Ca2+ and phospholipids. Structure of fibrinogen and its polymerization. Transglutaminase and cross-links. Prothrombin and its activation. Vitamin K and its role in the synthesis of gamma-carboxyglutamic acid (Gla). Anticoagulants and fibrinolysis. Carbohydrates. General properties and classification. Monosaccharides and glycosidic bond. Disaccharides, oligosaccharides and polysaccharides (starch, glycogen, cellulose). Reserve and structural carbohydrates. Heteropolysaccharides. Glycosaminoglycans and extracellular matrix. Glycoproteins. Proteoglycans. Lipids. General properties and classification. Saturated and unsaturated fatty acids. Reserve lipids. Triacylglycerols. Classification of membrane lipids. Phosphoglycerolipids and sphingolipids. Cholesterol and derivatives. Arachidonic acid and its derivatives. Composition and structure of biological membranes. Lipid and protein component of membranes. Membrane proteins: integral and peripheral. Passive and active transport mechanisms. Transporters (glucose transporters, chloro-bicarbonate transporter). Ion channels and pumps. Lipoproteins: structure, functions and recognition. Fat-soluble vitamins: A, D, E, K. The structure of nucleic acids - The structure of DNA. DNA stability: denaturation and renaturation (hybridization). The topological properties of DNA. Organization of chromosomes. The structure of RNA. Experimental methods for the separation of nucleic acids: electrophoresis and centrifugation. DNA-protein interaction Recombinant DNA technology - DNA amplification. Gene cloning. PCR and its applications in medicine. DNA sequencing. New biotechnology products. Recombinant proteins and biopharmaceuticals. Gene silencing (RNA interference). DNA editing (CRISPR-Cas9).

DL Nelson & MM Cox - Principi di Biochimica di Lehninger (ottava edizione) - Zanichelli D. Voet, JG Voet, CW Pratt - Fondamenti di Biochimica (quinta edizione) - Zanichelli