ECTS - University of Pisa
Il corso prevede conoscenze già acquisite di chimica generale, chimica organica e biochimica.
The course includes already acquired knowledge of general chemistry, organic chemistry and
biochemistry in order to adequately deal with the topics covered in the course.
La modalità di svolgimento dell'esame è orale.
The teacher assesses the knowledge acquired by the student during the oral test
Il corso offre allo studente l'opportunità di conoscere le moderne applicazioni della biochimica utlizzate
per la drug discovery, drug delivery, ingegneria proteica per applicazioni cliniche e produzione industriale di target
commerciali mediante l'utilizzo di bioreattori ed enzimi modificati.
The course offers to the student the opportunity to learn about the modern applications
of biochemistry used both in the field of basic research and in the pharmaceutical and food
industry. The student also learns about drug discovery, drug delivery and protein engineering.
Il docente valuta durante la prova d'esame se lo studente ha acquisito i concetti relativi alla applicazioni biochimiche presentate durante il corso.
The teacher assesses whether the student has acquired the concepts concerning the biochemical applications addressed during the course.
Il corso offre allo studente gli strumenti utili per comprendere come affrontare le problematiche relative alla stabilità chimico-fisica di una proteina,
a migliorare l'efficenza della catalisi di una proteina enzimatica, alla sua regolazione mediante effettori/inibitori e modulazione della stabilità del complesso proteina-ligando.
Questa conoscenza è utile per l'utilizzo di proteine e complessi proteina-ligando in ambito industriale e di diagnostica clinica.
The course offers the student useful tools to understand how to deal with problems
related to the maintenance or improvement of the chemical-physical state of a protein,
its regulation by effectors / inhibitors and the modulation of the stability of the
protein-ligand complex. This knowledge is useful for using proteins and protein-ligand
complexes in the industrial field and in clinical diagnostics.
In sede di esame lo studente dovrà dimostrare di conoscere gli aspetti metodologici utilizzati per lo studio di una proteina al fine di renderla utilizzabile in ambiti applicativi di diagnostica clinica e/o produzione commerciale.
The student will be asked during the examination which strategies are used in the biochemical field to regulate the catalytic capacities of an enzyme, the protein attitude in the interaction with its ligand and the most commonly used strategies for drug discovery and drug delivery
Lo studente deve essere a conoscenza di concetti acquisiti con il superamento dell'esame di Biochimica.
The student must be aware of concepts acquired with the passing of Biochemistry.
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not applicables
non richiesti
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Le lezioni frontali saranno accompagnate da slides che potranno essere successivamente scaricate sul sito apposito.
The lectures will be accompanied by slides that can be subsequently downloaded on the appropriate site.
Topics covered in the course and methods of examination. Ligand protein interactions.The association and dissociation constant of the protein-ligand complex.Presence of multiple non-equivalent binding sites. Evaluation of non-specific interactions. Graphical methods for Kd determination. Methods for determining the concentration of the protein-ligand complex. Overview of spectroscopic methods. Dialysis at equilibrium. Gel filtration chromatography, frontal chromatography. Use of enzymatic catalysis: general considerations. Synthesis of N-acyl-D-amino acids, S-derivatives of L-amino acids and alpha-keto acids. Phosphorylases for the synthesis of nucleoside and nucleotide derivatives. Immobilization of enzymes. Inosine synthesis marked on ribosidic portion. Enzyme immobilization methods: physical methods and chemical methods. Covalent immobilization; the activation phase of the support. Cross-linking. Effect of charges present on the support on the activity of the immobilized enzyme. Factors that can determine the loss of activity of an enzyme. The acquisition and maintenance of the correct folding: from the Anfinsen experiment to the molecular chaperones. The chaperone activity of the peptidylprolyl isomerase and the disulfide isomerase protein. The Hsp classes An example of Hsp60: E.coli's GroE system and its operation. Molecular chaperones: Examples of Hsp90 and Hsp70. The DnaK cycle and the role of Hsp90 in the maturation of steroid hormone receptors. The small heat shock protein: the example of alpha crystalline.
The alpha crystalline molecular chaperone in collaboration with other chaperones. Biotechnological application of molecular chaperones. The chaperone activity of the peptidylprolyl isomerase and the disulfide isomerase protein. The Hsp classes An example of Hsp60: E.coli's GroE system and its operation. Molecular chaperones: Examples of Hsp90 and Hsp70. The DnaK cycle and the role of Hsp90 in the maturation of steroid hormone receptors. The small heat shock protein: the example of alpha crystalline. The alpha crystalline molecular chaperone in collaboration with other chaperones. Biotechnological application of molecular chaperones Oxidative stress and antioxidant defenses of cells. Employent and independent enzyme detoxification mechanisms. Structure and function of glutathione Biosensors, general aspects. Biosensors used in clinical diagnostics. Biosensor for the determination of glucose, cholesterol, urea. Biosensors of first, second and third generation. Possibility of signal amplification. Biosensor for dependent metal enzymes Optical biosensors. Biosensor Lysine oxidase. Biosensors for pesticide determination. Biosensors containing immobilized algae. Biosensor for the evaluation of B.O.D. Brief affinity biosensors. Multienzymatic biosensor for the determination of fluid quality, introductory aspects. Multi-enzyme sensor for the evaluation of fluid quality. The site specific mutation: general aspects. Insulin: structure, biosynthesis and physiological role. Specific site mutations regarding insulin.Insulin Lys-Pro, Insulin AspB10 and examples of long-acting insulin. Specific site mutations: the example of glucose isomerase, glucosidase, subtilisin, phenylalanine, ammonia lyase. Biochemical foundations of Drug Discovery. Combinatorial chemistry techniques. Thioredoxin and thioredoxin reductase: structure and biological function. Synthetic and natural inhibitors of thioredoxin reductase: examples. Schistosomiasis. Thioredoxin Reductase inhibitors as drugs for therapy against Schistosoma diseases. Trypanosoma. Example of drugs against trypanosome diseases. The role of the Tripanotione. Research of Tripanotione Reductase inhibitors through in vitro and in silico methods Use of enzymes in the industrial field. Examples of enzymes involved in the degradation of food fibers, starch, phytic acid, polyphenols. Bioreactor for lactose hydrolysis. Use of hemicellulose for the production of biobutanol. Aldose Reductase type 2 (AKR1B1): general characteristics. Involvement of the polyol pathway in diabetic complications. Hyperosmotic model. Use of transgenic mouse lines for the study of diabetic cataracts. Involvement of AKR1B1 in the Naftalene-induced cataract. Definition of the reaction mechanism of the enzyme Aldoso Reductase by site-specific modification. Ruler of C298. Thiol-dependent modification of the Aldose reductase. Aloe-reductase modification from physiological disulphides. Effect of hyperbaric oxygen on the activity of the enzyme. Inactivation of the Aldose reductase by HNE and its oxidative modification by copper ion. Inhibitors of the Aldose Reductase type 2: classification. Tricyclic pyridazinones as selective inhibitors of the enzyme. Aloe reductase inhibitors as potential anti-cancer drugs: effect of topical treatment. Involvement of Aldose Reductase in the inflammatory response: correlation between Aldose reductase and activation of microglia cells, lens epithelial cells and macrophages. Role of Aldosa reductase in acrolein-mediated inflammation. Differential inhibitors of Aldose reductase. Role of Pyruvate Dehydrogenase Kinase in the onset of oncogenesis. Covalent inhibitors of Pyruvate Dehydrogenase Kinase. Role of Purines and Pyrimidines in the life cycle of an organism. Protective role played by purine and pyrimidine nucleosides against a human astrocytoma cell line subjected to ischemic insult.
Materiale didattico: slides presentate durante le lezioni frontali.
articoli scientifici
Teaching material: slides presented during the lectures. scientific articles
nessuna indicazione
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prova orale finale
oral exam
non presente
none
non applicabile
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