Syllabus:

1. Proteins (overview)

General properties, classification and importance for living organisms, structure and spatial arrangement of proteins with respect to their function , relationship between conformation and biological effects of proteins, denaturation.

2. Enzymes

Structure and mechanism of action of enzymes, effects of different factors on enzymatic activity, inhibition of catalytic activity of enzymes. Lineweaver-Burk plot, significance of Vmax and KM values, regulation of enzyme action in cells, regulatory enzymes and their control, specificity of enzymes, three-dimensional structures of enzymes, active site, isoenzymes, cofactors, coenzymes, nomenclature and classification of enzymes

3. Thermodynamics of the Living Systems

Concept of thermodynamics (TMD), laws of TMD, free energy, equilibrium, criterion of reaction spontaneity, coupled reactions, ATP – hydrolysis, biological significance, redox potential –importance, relationship between free energy and redox potential.

4. Metabolism of Carbohydrates

Carbohydrates - general information, formation of ring structures, glycolysis (chemical strategy of glycolysis, basic types of reactions in glycolysis, reactions of glycolysis), energy balance of glycolysis, the Pasteur effect, the Cori cycle. Regulation of glycolysis (alternative fates of glycolytic intermediates), gluconeogenesis (reactions and localization in the cell, regulation and energy balance), Entry of other carbohydrates into the glycolysis (monosaccharides -fructose, mannose , galactose, disaccharides), pentose phosphate pathway (importance, localization, reactions and regulation), key intermediates of metabolism, metabolic fates of pyruvate, pyruvate dehydrogenase complex. Glycogen Metabolism. Biosynthesis, breakdown and regulation, activation of kinases and phosphorylases, different metabolic pathway of glucose in various cells

5. Lipid Metabolism

Physiological role of lipids, fatty acids - the chemical nature , triacylglycerols – biosynthesis and degradation, control, utilization of fatty acids for energy production - activation, transport and reaction sequence for degradation, modifications of oxidative degradation, energy yield, regulation. Ketone bodies, structures, biosynthesis and control, peroxisomal oxidation of FA, reaction sequence for synthesis of palmitic acid and other fatty acids, comparison of degradation and biosynthesis of FA, regulation, metabolic interrelations of FA and carbohydrates in human body. Overview of membrane Lipids, Steroids and Cholesterol. Glycolipids, phospholipids, sphingolipids, structure, physiological role, biosynthesis and degradation, cholesterol: membrane component, precursor of bile salts and steroid hormones, synthesis of cholesterol, regulatory enzyme, regulation of biosynthesis, degradation of cholesterol: bile acids and steroid hormones, vitamin D.

6. Amino Acids Metabolism

Biological importance, energy yield, precursors of many biomolecules, ketogenic and glucogenic AA, metabolic reactions of amino- and carboxy-groups and their biological usefulness. Urea cycle – reaction sequence, control of urea cycle, location, metabolic degradation of AA carbon skeletons (individual AA). Reduction of N2, nitrogenase complex, assimilation NH4 to AA, nitrogenous derivatives of AA.

7. Citric Acid Cycle

Overview of the citric acid cycle, reactions and regulation of the citric acid cycle (CAC), amphibolic character of the CAC, anaplerotic sequences of the CAC, glyoxylate cycle.

8. Respiratory Chain and Oxidative Phosphorylation

Significance, localization, composition and function of complexes, cytochromes, inhibition of respiratory chain, Q cycle, respiratory control, chemiosmotic hypothesis, coupling of oxidation and phosphorylation, ATP synthesis and regulation.

9. Metabolism of Nucleotides

Synthesis of purine and pyrimidine ribonucleotides and deoxyribonucleotides, degradation of nucleotides, salvage pathways

10. Transport in Organism, Mitochondrion

Transport across membranes, nonmediated, passive and active transport, transport in mitochondrion

11. Heme and Hemoglobin

Synthesis of heme and its regulation, metabolism of heme to bile pigments, their significance and elimination.

12. Cell-Cell Signalling

Specific receptors mediating the cell signaling, G - proteins, cAMP a mediator of the action of many hormones, protein kinases, phospholipase C signalling pathway, receptors with enzyme function, lipophilic hormones and their cytosolic receptors. Structure and function of hormones, hormone systems and hormone hierarchy

13. Metabolic Interrelationship

Major metabolic pathways – localization, activity, control mechanisms, starve-feed cycle, well-fed state, early fasting state, fasting state, early refed state, glucose homeostasis, control mechanisms at all states, metabolic interrelationship in obesity, exercise and ethanol ingestion.

14. Basics of Xenobiochemistry

Introduction, history, cytochrome P450s, flavin-containing monooxygenases, reductases, conjugation pathways, induction and inhibition of drug metabolism, factors affecting drug metabolism, chiral aspects of drug metabolism, pharmacological and toxicological aspects of drug metabolism.

Practical Exercises

• Qualitative and quantitative determination of amino acids, Properties of protein solutions (isoelectric point, denaturation)

• Protein assays (Biuret reaction, Lowry method)

• Separation of low- and high-molecular weight samples (dialysis and gel filtration)

• Measurement of enzyme activity (activity, specific activity, temperature effect)

• Enzyme kinetics (basic kinetic parameters assessment – Michaelis constant, maximal velocity)

• Inhibition of enzymatic activity (assessment of inhibition concentration IC50, effect of inorganic ions)

Seminars

• Enzymes, enzyme kinetics, inhibition

• Metabolism of carbohydrates and their interrelationship, regulation

• Metabolism of lipids and amino acids, their interrelationship and regulation