Crystallization and preliminary X-ray analysis of the GST-fused human Bri3 N-terminal domain
- Department of Biochemistry, Queen’s University, Kingston, Ontario K7L 3N6 (Canada)
The crystallization of the polyproline-rich polypeptide from human Bri3 overexpressed as a GST-fusion protein in Escherichia coli is presented. Bri3 is a recently identified proline-rich transmembrane polypeptide up-regulated during TNF-mediated inflammation and immunity. The polyproline-rich N-terminal (residues 1–60) domain of Bri3 was affinity-purified to homogeneity as a glutathione-S-transferase (GST) fusion protein. Crystals were obtained in ∼3 d by the equilibrium vapour-diffusion method from a solution containing 1.5–2.2 M ammonium sulfate and 0.1 M bis-tris pH 6.0. The crystals belong to space group P4{sub 3}2{sub 1}2, with unit-cell parameters a = b = 91.66, c = 57.53 Å. An X-ray data set was collected to 1.6 Å resolution using synchrotron radiation, with an R{sub sym} of 0.058 and a completeness of 95.3%. There is one molecule of the fusion protein in the asymmetric unit, which corresponds to ∼35% solvent content.
- OSTI ID:
- 22356081
- Journal Information:
- Acta Crystallographica. Section F, Vol. 61, Issue Pt 1; Other Information: PMCID: PMC1952390; PMID: 16508092; PUBLISHER-ID: bw5068; OAI: oai:pubmedcentral.nih.gov:1952390; Copyright (c) International Union of Crystallography 2005; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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