DARTS NEW PROTEIN CRYSTALLISATION SERVICE

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DARTS NEW PROTEIN CRYSTALLISATION SERVICE – HOW IT WORKS
01 October 2003 - CCLRC Daresbury Laboratory

Pharmaceutical companies are in the business of producing drugs designed to interact with specific protein targets. The drug/target interaction is first tested in order to elicit an effect, either boosting the protein's biological activity or reducing it. To fine tune the interaction, further detailed information is usually required, which often means structure determination of the protein coupled to its target molecule. DARTS is well disposed to offer a complete protein crystallisation service to satisfy your needs.

As the customer, you may want to provide DARTS with a 'ready to use' concentrated sample i.e. stable at 5 - 10 mgml-1, monodisperse and with known characteristic profiles. DARTS can pre-check these characteristics after shipping either by SDS-PAGE or UV-VIS in addition to DLS measurements to assess monodispersity. DLS can also be applied to determine the most suitable temperature and/or pH for producing crystals. If stability is an issue during transportation, DARTS can provide a range of final purification steps and concentration of dilute protein samples.

DARTS uses commercial crystallisation screens to probe 100-200 different conditions at two different temperatures. The initial screen requires 500 µl of concentrated protein. The screen is then refined until diffraction quality crystals are obtained. Most common crystallisation problems arise due to poorly ordered protein crystals such as those with glycosylation sites containing flexible sugar molecules or membrane proteins with a high solvent content and containing detergent molecules. DARTS can also try crystallisation conditions that have already been reported in the literature for similar target proteins.

DARTS staff have been successful in crystallising a number of globular and membrane proteins e.g. light-harvesting complexes, plant lectins and bacterial phosphatases. Some of these crystals diffracted to atomic resolution (better than 1.2 Å). In addition, staff are technically adept in data acquisition, structure solution and refinement. This powerhouse of expertise can be made available for the most pressing needs of today's pharmaceutical industries. We are confident that DARTS can provide a service to suit your needs.

http://www.darts.ac.uk

About: CCLRC Daresbury Laboratory
DARTS is a unique service offering solutions to materials characterisation problems that are unattainable in the conventional laboratory. This is possible because it makes use of the Synchrotron Radiation Source (SRS). Advantages of using the SRS include:

• The ability to study small or weakly diffracting samples. Improved resolution, enabling structures intractable in the home laboratory to be solved.

• The ability to select the optimum wavelength for a particular application.

• Additional specialised techniques only possible using synchrotron radiation can be exploited.

The DARTS team's ability to access and combine a range of complementary techniques provides information to help manufacturers control processing conditions and modify manufacturing techniques to improve a wide range of products. Even the everyday crisp packet has benefited from DARTS leading edge technology.

Applications can cover materials from agrochemicals, pigments, polypeptides, microporous materials, organometallics, catalyst materials, minerals and samples from processing plants.

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