Enzymes are proteins which function as biological catalysts, playing an important role in nearly all processes taking place in living organisms. As a result, studies of these biological molecules and the reactions they catalyse is of interest, especially when considering the development of novel therapeutics against diseases such as cancer. A number of drugs are already available which inhibit enzyme activity. Development of new inhibitors requires an understanding of the way an enzyme binds and then process the natural substrate.
In this webinar we will consider how Isothermal titration Calorimetry (ITC) can be applied to understand enzyme inhibition and activation. ITC was originally designed for studying host/guest binding interactions but is gaining popularity as general enzyme assay. To characterize enzyme activity, ITC measures the heat released or absorbed by catalysis in real time, following the rapid mixing of enzyme and substrate solutions. Since most chemical reactions are either exothermic or endothermic, ITC can be applied to virtually any enzyme/substrate pair, without the need to design customized reporter molecules, to couple the reaction to additional enzymes, or to perform any post-reaction separation. In addition, ITC experiments can be performed under dilute, physiological solution conditions, even with opaque samples and require far less enzyme than traditional ITC binding experiments.
Basic concepts in ITC enzyme assays will be introduced together with recent developments from our lab, followed by some case studies that illustrate the capabilities of the technique. The results we present help illustrate the broad applicability of ITC to better understanding enzyme inhibition.
September 24 2019 - September 24 2019
10:30 - 11:30
(GMT-05:00) Eastern [US & Canada]
Webinar - Live
Anthony Mittermaier Ph.D. – Associate Professor McGill University Department of Chemistry
Anthony Mittermaier obtained his B.Sc. in biophysics at the University of Guelph in 1996 and his Ph.D. in biochemistry at the University of Toronto under the supervision of Lewis Kay and Julie Forman-Kay in 2003. After a short post-doc under the supervision of Lewis Kay and Regis Pomes at the Hospital for Sick Children in Toronto, he joined the Department of Chemistry at McGill University in 2005, and was promoted to Associate Professor in 2011. His research employs a combination of biophysical techniques, particularly calorimetry and NMR spectroscopy, to address fundamental questions regarding the molecular basis of protein and nucleic acid function and drug design. In 2012 he was awarded the Agilent Early Career Professor Award for his advances in combining NMR with calorimetric techniques. More recently, his laboratory has begun to focus on using ITC to measure enzyme kinetics and thermal hysteresis experiments to measure supramolecular assembly.
- How to directly follow enzyme activity with ITC without a need to design customized reporter molecules, to couple the reaction to additional enzymes, or to perform any post-reaction separation.
- How ITC can be applied to understanding enzyme inhibition and/or activation.
- General concepts associated with the development of ITC enzyme assays
- Scientists in academia and industry working with enzymatic assays and characterization of enzyme activity, mechanism and inhibition.
- Drug developers engaged in hit identification and validation for drug products targeting enzyme activity