Biochemistry

General skills The purpose of this course is to provide the students with the fundamental intellectual tools required to carry out measurements, analyses and interpretation of enzyme kinetics. At the end of the course, the students will be confident with the elementary concepts of chemical kinetics, will understand the theoretical basis of steady state and rapid equilibrium enzyme kinetics, and will be able to derive the relevant rate equations. They will also be familiar with the practical aspects of enzyme kinetics, such as enzyme assay methods and the use of computer software to analyse kinetics data. The understanding and analytical skills of the students will include enzyme reactions with more than one substrate, enzyme inhibition and activation. Concerning the transient phase of enzyme reactions, students will understand the theoretical basis of rapid kinetics and the main experimental techniques used for their measurement; they will also be able to analyse and interpret rapid kinetics. Specific skills At the end of the course the student will acquire a) knowledge and understanding of:  theoretical principles of single substrate and multisubstrate steady-state and rapid equilibrium kinetics;  practical aspects linked to the measurement and analysis of enzyme kinetics, which include the various enzyme assay methods but also the correct determination of reaction velocity from a progress curve, the strategies for determination of kinetic and equilibrium constants using graphical methods and computer software, the determination of active site concentration and the statistical analysis of enzyme kinetics data;  the effect of pH and temperature on enzyme activity and kinetic parameters;  the main types of enzyme inhibition and activation, including complex mechanisms deriving from allosteric and non-allosteric interactions with small molecules; students will also know and understand irreversible, mechanism-based inhibition; in this context, students will appreciate how enzymes are regulated in the metabolism and how enzyme inhibition can be used as a means of control of cell metabolism in medicine;  theoretical principles of pre-steady state kinetics and also the main experimental techniques and instruments used for the measurement of rapid kinetics, including continuous-flow, stopped-flow, laser and relaxation methods. b) ability to apply knowledge and understanding:  apply the principles of steady-state and rapid equilibrium kinetics to derive equations describing the behaviour of single substrate and multisubstrate reactions;  apply these equations to the analysis of kinetic data, using graphical methods and computer software, in order to estimate kinetic and equilibrium parameters;  apply knowledge on practical aspects of enzyme kinetics to carry out correct experimental measurements and analysis of kinetic data;  derive rate equations that describe simple and complex activation and inhibition systems, and apply such equations to analyse kinetics data and estimate kinetic and equilibrium constants;  apply knowledge on the transient phase of enzyme reactions to derive rate equations describing pre-steady state kinetics; apply these equation, using computer software, to estimate kinetic parameters.  at the end of the course, students will understand how enzyme kinetics data can be used in the study of structure-function relationships of proteins. c) autonomy of judgment:  for a specific enzyme kinetics study, choose the most appropriate experimental set up and measurement method;  judge the goodness of the experimental results in terms of reproducibility and replicability;  recognize the main features of the system under study, so as to derive or identify in the literature the equations required for data analysis; use these equations to analyse the experimental data in quantitative terms, using appropriate computer software;  interpret the outcome of kinetic analyses, so as to propose the most appropriate theoretical model describing the behaviour of the experimental system under study. d) communication skills:  produce PowerPoint presentations with experimental data, equations and graphs;  for a specific enzyme kinetics system, explain the analysis of experimental data and their interpretation, present a theoretical model that explains experimental data;  trace the chosen theoretical model back to structure-function relationships. e) learning ability:  autonomously face and solve novel enzyme kinetics problems;  independently continue and make progress in the study of enzyme kinetics;  learn independently, or with limited supervision, how to apply the skills acquired in this course to the experimental laboratory work.