NMR is a versatile tool to study the biophysical characteristics of enzymes. In my research, I use various NMR experiments to investigate the enzyme isocitrate lyase from Mycobacterium tuberculosis. This enzyme was found to be essential to the survival and pathogenesis of the bacteria, but currently our understanding about its catalytic mechanism and modulation are limited. Thus, targeting the regulation of this enzyme may shed light on a better solution in eradicating this widespread pathogen.
One of the methods of enzyme regulation is through protein-metabolite interactions. Water ligand-observed via gradient spectroscopy (waterLOGSY) was used to study the binding of metabolites to isocitrate lyase. WaterLOGSY is a high-throughput technique that gives clear indication of binding, making it useful for the screening of metabolites. 1H-NMR was utilised in investigating the kinetics of isocitrate lyase, especially in cases when the classic UV-visible absorbance-based assays were unsuitable for analysis. Finally, the dynamics of domain dimerisation in isocitrate lyase was examined via protein NMR. Static structures from X-ray crystallography, coupled with dynamic information from protein NMR, can provide a full picture of this domain dimerisation. Overall, this study highlighted the different NMR techniques that can be used in the analysis of enzymes, and it showed that NMR is able to enhance the capabilities of other biophysical techniques when certain circumstances preclude these techniques from being used or prevent them from providing the complete explanation to a phenomenon.