Crystallization and preliminary X-ray diffraction analysis of vaccinia virus H1L phosphatase
- Departamento de Química Física y Analítica, Universidad de Oviedo, Julián Clavería 8, 33006 Oviedo (Spain)
- Structural Biology Laboratory, London Research Institute, Cancer Research UK, 44 Lincoln’s Inn Fields, London WC2A 3PX (United Kingdom)
- ESRF, 6 Rue Jules Horowitz, 38043 Grenoble CEDEX 9 (France)
- Cell Motility Laboratory, London Research Institute, Cancer Research UK, 44 Lincoln’s Inn Fields, London WC2A 3PX (United Kingdom)
A catalytically inactive mutant of the dual-specificity phosphatase H1L from vaccinia virus was expressed recombinantly, purified and crystallized by the microbatch method. The crystals belong to the tetragonal space group P422 and diffraction data were collected to 2.1 Å resolution using a synchrotron-radiation source. Attempts to derivatize these crystals with xenon gas lead to a space-group change to I422 with a smaller unit cell and a diffraction limit of 3.0 Å. The cysteine-based protein phosphatase H1L was the first reported dual-specificity protein phosphatase. H1L is encapsidated within the vaccinia virus and is required for successful host infection and for the production of viable vaccinia progeny. H1L has therefore been proposed as a target candidate for antiviral compounds. Recombinant H1L has been expressed in a catalytically inactive form using an Escherichia coli host, leading to purification and crystallization by the microbatch method. The crystals diffract to 2.1 Å resolution using synchrotron radiation. These crystals belong to space group P422, with unit-cell parameters a = b = 98.31, c = 169.15 Å, and are likely to contain four molecules in the asymmetric unit. A sulfur SAD data set was collected to 2.8 Å resolution on beamline BM14 at the ESRF to facilitate structure determination. Attempts to derivatize these crystals with xenon gas changed the space group to I422, with unit-cell parameters a = b = 63.28, c = 169.68 Å and a single molecule in the asymmetric unit. The relationship between these two crystal forms is discussed.
- OSTI ID:
- 22360483
- Journal Information:
- Acta Crystallographica. Section F, Vol. 64, Issue Pt 3; Other Information: PMCID: PMC2374156; PMID: 18323605; PUBLISHER-ID: gj5036; OAI: oai:pubmedcentral.nih.gov:2374156; Copyright (c) International Union of Crystallography 2008; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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