SUBJECT

Enzymology

lecture

master

Faculty of Science

Biology MSc

2

Semesters 1-4

Autumn/Spring semester

1. Enzymes in vivo: proteins and ribozymes. Moonlighting enzymes. Artificial and applied enzymes

2. Enzyme kinetics. The progress curve. The dependence of initial velocity on substrate concentration: first-, non-integral- and zero-order parts of the substrate saturation curve. The specificity constant, the imiting velocity and the Michaelis constant.

3. Determination of steady-state parameters by linear and direct plots. Analysis of the progress curve.

4. Effects on environment on enzyme activitz: pH-dependence. Experimental conditions, parameters. Irrev ersible and reversible inhibition: the competitive(specific) and the catalytic one.

5. Analysis of temperature dependence. From kinetic parameters to molecular mechanism.

6. Enzyme catalysis and specificity: the protein structural ground. Levels of protein structure and their hierarchy. Amino acid side chains and elementary catalytic factors. Redox coenzymes.

7. Stability and flexibility of protein structure. Concerted catalysis and induced-fit. Molecular recognition: substrate specificity. Active centre and -site.

8. Stereochemistry of catalysis. Transition state stabilisation, proximity and orbital steering. Reaction specificity: enzyme classes.

9. Control of enzyme activity by irreversible and reversible covalent modification: limited proteolysis, protein kinases and phosphatases.

10. Metabolite control of enzyme activity: allostery and cooperativity. Protein complexes.

11. Anatomy of chymotrypsin catalysis Chemical modifications: chymotrypsinogen activation, identification of the serine-histidine diad. Kinetic studies: acylenzyme and tetrahedral intermediate.

12. X-ray crystallography: stereochemistry of activation, catalytic triad, oxyanion hole.

13. Technological enzymes and protein engineering Enzymes in food- and pharmaceutical industries, in analytical chemistry and in the household. Enzymes as tools and targets of protein engineering.

14. New perspectives: thermophile enzymes, abzymes, molecular imprinting.

• Tim Bugg: An Introduction to Enzyme and Coenzyme Chemistry

• Blackwell Sci. Ltd, 1997

• Cornish-Bowden, A.: Fundamentals of Enzyme Kinetics, Portland Press, 1995, ISBN 1 85578 072 0

• Selected overheads of lecture