Synchrotron crystallography team
Figure 1: Coffee beans contain large quantities of beneficial compounds such as 5-CQA and 3,5-diCQA that are predominantly synthesised by HQT and HCT respectively. The lower left panel shows a chromatogram illustrating the synthesis of 3,5-diCQA catalysed by a HCT mutant. The central inset shows the best docking result of 3,5-diCQA in HCT.
Figure 2: Diffraction Viewer (dViewer) software is currently under development for the visualisation of new generation pixel detector images
The McCarthy team works on the design, construction and operation of MX and BioSAXS beamlines and studies proteins involved in neuron development.
Previous and current research
Our team works in close collaboration with the Structural Biology Group of the European Synchrotron Radiation Facility (ESRF) in the design, construction and operation of macromolecular crystallography (MX) and biological X-ray scattering (BioSAXS) beamlines. We are currently responsible for two macromolecular beamlines, ID14-4 and ID23-2, the BioSAXS beamline at BM29, as well as the commissioning of MASSIF1 on ID30. The team also manages the operation of BM14, which is run as a partnership with the ESRF and the Indian government. The structural biology beamlines at the ESRF continue to perform, resulting in the deposition of more than 898 structures in the PDB last year. We work in close collaboration with the Cipriani team to develop hardware and we have successfully commissioned a plate holder for the in situ screening of crystals on BM14, which will soon be available to external users. We are also developing novel methodologies for advanced sample screening and data collection possibilities. A recent example is the use of workflows in the design and implementation of complex experiments in MX.
We study proteins involved in neuronal development, particularly the Slit-Robo signalling complex, and proteins involved in the synthesis of plant secondary metabolites (figure 1). Meanwhile, the BM14 group is actively involved in structural studies of proteins involved in Toxoplasma gondii epigenetic machinery in collaboration with Mohamed-Ali Hakimi (Grenoble Medical University).
Future projects and goals
This year will be exciting, with the completion of the next phase of the UPBL10 project with our ESRF colleagues, the first user operation of the new MASSIF (Massively Automated Sample Selection Integrated Facility) suite of MX-beamlines on ID30A. This ambitious project is part of the ESRF upgrade program and will ensure that European users will have continued access to state-of-the-art structural biology beamlines for the next decade. On BM14, we will optimise the in situ screening of crystals in plates and install a new sample changer design for commissioning. The team will continue to develop automated data screening, collection and analysis schemes, phasing methods using UV induced damage, as well as a new diffraction image viewer software developed using Eclipse-RCP technology and as part of BioStruct-X and DAWN collaborations (figure 2). We will further develop the highly automated BioSAXS beamline in collaboration with the ESRF, the Cipriani team and colleagues at EMBL Hamburg, including the full integration of an online HPLC system with additional biophysical characterisation features. We hope that all our combined efforts will push the boundaries of structural biology to better understand the functions of more complex biological systems.
In our laboratory, we will continue research on the Slit-Robo signalling complex by trying to decipher exactly how Slit activates Robo on the cell surface. We will also extend work on phosphoryl transfer into human kinase signal cascades. The BM14 group will over-express several Toxoplasma proteins in E. Coli to be structurally and biochemically characterised using the PSB facilities. Lastly, the initial work on a novel dequibiquitinase will be continued.