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Enzyme dynamics: NMR spectroscopy and kinetics (EDNMR)

Course 2018-2019

Core course in MSc Chemistry - Chemical Biology, core course in MSc Life Science and Technology

Description

Proteins carry out most functions in the cell. These molecules are not rigid, but are mobile on a wide range of timescales. The course aims to convey the concept of protein dynamics in relation to biological function. The focus is on enzyme catalysis and two techniques will be discussed in detail: nuclear magnetic resonance (NMR) spectroscopy to study protein mobility and kinetic measurements to characterise enzyme activity. NMR is a versatile analytical technique with many applications. We will discuss the fundamentals of NMR (chemical shift, dipolar coupling, J-coupling, relaxation), as well as multidimensional NMR and relaxation experiments to characterize molecular mobility on nanosecond and millisecond timescale. It provides a basis in NMR spectroscopy that is of use for any chemist. The study of kinetics will comprise steady state and pre-steady state theory and methods to measure enzyme characteristics such as k(cat) and K(M), as well as inhibitor kinetics. The theory will be placed into context by studying several papers from the literature.

Methods used

Lectures, exercises, discussions and simulations. We will work with a flipped class-room approach for some aspects of the course.
Learning goals:
• the student understands the fundamentals of NMR spectroscopy and can explain the concepts of chemical shift, dipolar coupling, J-coupling, relaxation
• the student understands how NMR spectroscopy can be used to study protein dynamics on different timescales and can critically discuss possibilities and limitations
• the student understands and can apply the theory of steady state and pre-steady state kinetics for the characterisation of enzyme-catalysed reactions in calculations and simulations
• the student can explain how kinetic experiments are performed and analysed, both for steady state and stopped-flow measurements
• the student understands the concept of dynamics in enzymology and can apply this knowledge to explain results described in the literature as well as to address questions concerning new systems

Evaluation

Exercises and an on-line test need to be completed and handed-in to be allowed to participate in the written exam after the course.

Examination

Written exam with admission requirement.

Knowledge at entry

Good understanding of protein structure (primary – quaternary structure, amino acids), basics of Michaelis Menten kinetics, concept of Boltzmann equilibrium.

Registration

Register for this course via uSis