A high-throughput crystallisation facility for protein structure determination

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The aim of this application is to provide a robotic nanolitre protein crystallisation facility for the Centre for Structural Biology (CSB) at Imperial College London. The CSB is a college centre with affiliated groups from three faculties and provides both expertise and managed core facilities in all major techniques of macromolecular structure determination: X-ray crystallography, nuclear magnetic resonance spectroscopy, and electron microscopy. The research carried out by CSB members (with substantial BBSRC funding) is of the highest international standard and embraces a wide spectrum of topics; membrane proteins; mechanisms of DNA replication, transcription and repair; bacterial gene regulation; enzymes and molecular machines; mechanisms microbial pathogenicity; structural basis for human diseases and drug binding; and method development. X-ray crystallography is used as the major technique for structure determination by 10 research groups and critically contributes to the research of many other groups, thanks to a number of well-established multidisciplinary collaborations. State-of-the-art equipment exists for protein characterisation (funded by BBSRC-REI 2004) and for X-ray diffraction data collection, but the progress of many structural biology research projects is severely hampered by the lack of a robust high-throughput facility for robotic nanolitre protein crystallisation. Therefore, we request the following instrumentation: (1) a robotic liquid handler to fill 96-well plates with commercial or customised crystallisation solutions; (2) a crystallisation robot to set up nanolitre volume crystallisation experiments. These instruments will be managed by the CSB and provide a central crystallisation facility serving over 100 users. This facility will underpin and enhance many BBSRC-supported grants, as well as other research projects. The instruments will complement existing equipment and significantly improve the research infrastructure of a large number of highly productive groups within the CSB. Crystallisation facilities similar to the one envisaged in our application have been operating successfully at several other top UK universities; we strongly believe that the requested instrumentation will be essential for the CSB at Imperial College London to maintain its internationally leading position. We believe that the requested instruments offer good value for money, as they will very significantly reduce the expenses of protein production and crystallisation of a large number of users. Equally important, robotic crystallisation will be essential to make progress in the emerging areas of system-wide structural biology. In summary, we believe that a well-managed crystallisation facility within the CSB will greatly enhance the research infrastructure of a large number of active researchers, complement existing multi-user equipment, and critically contribute to the training of our students and postdocs.