Research Facilities in X-ray Crystallography
A suite of protein crystallography laboratories, featuring robotic crystallization, dedicated crystal storage, automated crystal viewing, state of the art X-ray data collection facilities, high speed data processing and molecular graphics complements our fully specified molecular biology and protein purification laboratory, where the biological samples to be studied are prepared. |
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Figure 1 [left] Overexpression and preparation of pure protein, [right] Crystallisation robot. |
| Having first identified, cloned, overexpressed and purified the target protein of interest, a fully automatic Matrix Hydra II PlusOne crystallization robot is used to test many hundreds of possible crystallization conditions. Trials are stored in a dedicated, temperature stable crystal hotel, and checked using a semi-automatic plate reader for possible crystal hits. |
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Figure 2 [left] Crystal trials stored in a temperature stable crystal hotel and [right] Evaluating potential crystals on semi-automatic plate reader. |
| Crystals are screened and data collected on the in-house X-ray facility. X-rays are produced using a Rigaku MicroMax 007 micro-focus copper rotating anode generator running at 40kV, 20mA. Two data collection stations are mounted on the generator. For each station the X-rays are focussed onto the crystal using confocal multilayer optics. One station uses Osmic Varimax MaxFlux optics to optimise the beam intensity at the crystal, with the other station optimised for crystals with long unit cells using the Osmic Varimax HighRes optics. For each data collection station, the crystal is cooled to 100K with an Oxford Cryosystems Cryostream nitrogen cooler, and data accumulated using a MarResearch MAR345 image plate detector system. Data are processed, structures determined and molecular structures viewed using a cluster of Unix, Linux and OSX computers. |
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Figure 3 [left] In-house X-ray Facility, [right] Experimental Station |
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Funded : through grants from BBSRC, EPSRC, Wellcome Trust, The Royal Society, Wolfson Foundation, EU and NEDO |