Current Research Interests
I am currently involved in work on the following areas:
- Development of applied methods using in-situ plate screening
- Beamline automation and instrumentation
- Structural biology (protein-ligand interactions)
In-situ plate screening
We are looking at developing applied methods in crystallography, by taking advantage of the high-throughput in-situ plate screening setup of the beamline (10 to 15 conditions can be screened per hour). As such, we have implemented a simple and relatively fast dehydration procedure that allows, in a single experiment, a comprehensive screening of the dehydration effect on protein crystals with X-ray feedback. In collaboration with Petra Lukacik, we have applied this method to a membrane-associated protein and monitored the effect of the dehydration on the crystals by following the shrinkage of the cell parameters. In this case, the result of dehydration also led to an improvement of the crystal’s diffraction from 2.9 Å resolution to 2.45 Å. We are further evaluating this method for membrane protein crystals in collaboration with Isabel De Moraes from the Membrane Protein Laboratory (MPL).
I would welcome enquiries from interested users for further information on this method, as well as potential collaborations.
In collaboration with Irelec and I24 beamline, we have developed an end-effector compatible with the CATS robot (Irelec) and the unipuck sample holder (16 samples per puck) for a high capacity and faster sample exchange system. The prototype is currently running at the beamline with an exchange time reduced from 150 seconds to 37 seconds. We are also looking into easing the transition from the cryogenic experiments using the robot in the cryotong mode to the plate screening mode at room temperature. This improvement would enhance the flexibility of the beamline allowing users to operate two modes within their allocated 24-hours beamtime.
My previous experience in the pharmaceutical industry involved work on the characterisation of protein-small molecule complexes for the design of drug molecules using primarily protein crystallography but also other biophysical methods. X-ray crystallography is a very powerful tool in drug discovery, enabling a rational design based on the knowledge of the atomic details of protein-ligand interactions. Methionine aminopeptidase (MetAP) is a metalloprotease that removes the N-terminal methionine during protein synthesis and is essential for bacterial survival. Mycobacterium tuberculosis expresses two methionine aminopeptidases, MtMetAP1a and MtMetAP1c and both are potential targets to develop novel antitubercular therapeutics. There is accumulating biochemical evidence that both forms have large differences in their active site and therefore could potentially lead to different classes of inhibitors. Several constructs of both MtMetAP1a and MtMetAP1c are currently being investigated as part of the protein 100 project at the Oxford Protein Production Factory (OPPF), to understand the selectivity of both forms, and might be the foundation for structure-based small molecule design projects.
Isabel De Moraes (Membrane Protein Laboratory @ Diamond Light Source, UK)
Matthieu Privat De Fortuné and Kenji Laroche (Irelec, Grenoble, France)
Ray Owen, Louise Bird, Jo Nettleship, Yamini Reddivari (Oxford Protein Production Factory, Research complex @ Harwell science and innovation campus, UK)
Jose Brandao-Neto, Pierre Aller, Petra Lukacik (Diamond Light Source, UK)