McCarthy Team
Synchrotron Crystallography Team
Figure 1: View of the highly automated ID14-4 experimental hutch
Figure 2: Structure of the Slit2 dimerisation domain. Slit2 D4 N- and C-terminal caps are in purple and blue, respectively, the LRRs 1-5 are in orange and the disulphide bridges are in yellow.
Previous and current research
The team works in close collaboration with the structural biology group of the ESRF in the design, construction and operation of synchrotron radiation beamlines. We are currently responsible for two macromolecular beamlines, ID14-4 (figure 1) and ID23-2, as well as the BioSAXS beamline at ID14-3. The team also manages the operation of BM14, which is run as a partnership with the ESRF and Indian government. BM14 is now welcoming users from India as well as EMBL member states. All the structural biology beamlines at the ESRF are performing well and a particular highlight is the continued use of the beamlines by the groups of Venki Ramakrishnan and Ada Yonath, who were awarded the 2009 Nobel prize in Chemistry with Thomas Steitz for their ‘studies on the structure and function of the ribosome’. We also work in close collaboration with the diffraction instrumentation team to develop hardware, software and novel methodologies for sample handling and data collection possibilities. Recent examples include the high throughput BioSAXS liquid handling robot recently installed on ID14-3 and soft ware for optimised MX data collection using crystal reorientation strategies.
The team also studies proteins involved in neuronal development. We are particularly interested in the Slit- Robo signalling complex that is essential for the normal development of the central nervous system. This signalling system has also been implicated in heart morphogenesis, angiogenesis and tumour metastasis. With part funding from SPINE2Complexes we have determined a number of structures from this system that maybe important for the development of novel cancer therapeutics (figure 2). We are also interested in understanding the molecular mechanism of proteins involved in the biosynthesis of plant secondary metabolites. To this end, we have already published the structures of two enzymes involved in caffeine biosynthesis. These studies suggest it may be possible to generate a single protein capable of producing caffeine in plants. Such a possibility, when coupled with caffeine’s ability to act as a natural pesticide, could enable new ecologically friendly and pest resistant plants to be created.
Future projects and goals
This year we will commence work on the UPBL10 project with our ESRF colleagues. 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 10 years. This project will commence with the construction of a new bioSAXS beamline on BM29, which should welcome users later this year. A major upgrade of the X-ray optics for BM14 will also be completed this year, enabling a smaller and brighter X-ray beam for improved performance. The team will continue to develop novel data collection schemes using the MK3 for challenging structural biology projects and the integration of X-ray tomography methods in MX. On ID23-2 we plan to continue to develop specialised methods for the handling and collection of optimal data from ever smaller crystals. On ID14-3 our team, in collaboration with the diffraction instrumentation team, the ESRF, and EMBL Hamburg, will continue to be actively involved in the provision of a highly automated BioSAXS beamline. We hope that all our combined efforts will push the boundaries of structural biology to better understand the biological functions of more complex biological systems.
In the laboratory we will continue our research on the Slit-Robo complex by trying to decipher how exactly Slit activates Robo on the cell surface. We plan to tackle this by studying larger fragments of Robo and Slit and using complementary methods to MX where necessary. In collaboration with Néstle Research, France, we plan to try and expand our current research on secondary metabolic pathways in coffee.