CELLULAR BIOCHEMISTRY AND PROTEOMICS
A solid background in Citology, Biochemistry and Molecular Biology.
The examination, oral only, consists of two questions. The first concerns the organization of the exposure of a subject of the course, without specific guidance, in order to evaluate the autonomy to make judgments, as well as knowledge and understanding. The second question is more specific and is focused on the analysis of specific problems within a single subject, to assess the ability to apply knowledge and understanding.
The main objectives of the course are:
-to understand the mechanisms of biochemical regulation of cell function.
-to acquire knowledge about post-genomic methods in cellular biochemistry.
CELLULAR BIOCHEMISTRY- 5 CFU
• Introduction to cellular biochemistry. Course objectives and examination modes
• Protein homeostasis. Protein synthesis. Post-translational translocation. Signal sequences. Intracellular protein trafficking. Quality control. UPS degradation. The secretory pathway. Molecular mechanisms of vesicular trafficking. Coating proteins and clathrin. Proteolytic processing of secreted proteins. Receptor-mediated endocytosis. Synaptic secretion. Autophagy.
• Post-translational modifications. The ability of particular tags to regulate signaling and protein turnover, and to exert dynamic control over protein function in diverse cell biological contexts.
• Membranes and transport. Chemical components of membranes. Peripheral and integral proteins. Lipid rafts. Mechanisms of membrane fusion. Transport processes. Cooperativity and allostery in membrane transporters.
• Cytoskeleton. Three kinds of filaments building the cytoskeleton. Actin filaments: polymerization, dynamics, myosins, cell motility. Intermediate filaments: structure and mechanics. Intermediate filament associated proteins. Microtubules: molecular organization and polarization. Proteins regulating microtubule dynamics. Microtubule organization centers. Motor proteins: Kinesins and dyneins. The mitotic apparatus.
• Integration of cells into tissues. Adhesive junctions. Adhesive proteins. Biochemistry of cell junctions: adherent junctions, desmosomes, hemidesmosomes, tight junctions, gap junctions. The basal lamina. Proteins of the extracellular matrix.
• The mitochondrion. Cell energetics. Mitochondrial biochemistry. Transport towards and across the inner membrane. Glucose and fatty acid metabolism. Biochemistry of the electron transport chain. Calcium ion homeostasis. Apoptosis regulation. Mitophagy. Organelles biosynthesis. Mitochondrial dynamics.
• Biosignaling. Characteristics of a transduction system. Molecular mechanisms of signal recognition. Ligand gated ion channels (e.g., the nicotinic receptor, neuronal signaling). Receptors with intrinsic enzyme activity (e.g., glycogen synthesis). G-protein-coupled receptors (e.g., taste and smell transduction). Receptors linked to soluble kinases (e.g., the erythropoietin receptor). Guanylate cyclase activity and NO-synthase activation. Adhesion receptors. Intracellular receptors.
• Respose to stress. Oxidative stress. Reactive oxygen species. Enzymatic control of ROS homeostasis. Iron homeostasis. Lipid oxidation. Protein oxidation. Unfolded protein response.
• Hormone regulation and metabolic control. Molecular aspects of hormone action. Peptide hormones. Catecholamines. Eicosanoids. Steroid hormones. Vitamin D. Retinoids. Thyroid hormones. The neuroendocrine system. Metabolic specialization of tissues: liver, muscle, brain and pancreas. The hypothalamus.
PROTEOMICS- 2 CFU
• Post-genomic methods in cellular biochemistry. The principles of the “a posteriori” hypothesis
• Integrated Omics: Transcriptomics, Proteomics, Metabolomics.
• Proteomics approaches and technologies: In-gel and off-gel, quantitative analysis, statistical approaches, databases for protein identification.
• Biochemical strategies to reduce sample complexity.
• Beyond proteomics. MALDI imaging, MALDI profiling, Metabolomics.
All the presentations, reviews, research articles and relevant material will be loaded on the course e-learning platform.
Only lectures given by the teacher in the presence of students. Seminars by experts in the field will be considered for particular topics.