Crystallization and X-ray diffraction analysis of the complement component-3 (C3) inhibitory domain of Efb from Staphylococcus aureus
- Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri-Kansas City (United States)
- Division of Rheumatology, Department of Medicine, The Johns Hopkins University School of Medicine (United States)
The crystallization and results of multiwavelength anomalous diffraction studies of a recombinant C3-inhibitory fragment of Efb from S. aureus are reported. The extracellular fibrinogen-binding protein (Efb) of Staphylococcus aureus is a multifunctional virulence factor capable of potent inhibition of complement component-3 (C3) activity in addition to its previously described fibrinogen-binding properties. A truncated recombinant form of Efb (Efb-C) that binds C3 has been overexpressed and purified and has been crystallized using the hanging-drop vapor-diffusion technique. Crystals of native Efb-C grew in the tetragonal space group P4{sub 3} (unit-cell parameters a = b = 59.53, c = 46.63 Å) with two molecules in the asymmetric unit and diffracted well beyond 1.25 Å limiting Bragg spacing. To facilitate de novo phasing of the Efb-C crystals, two independent site-directed mutants were engineered in which either residue Ile112 or Val140 was replaced with methionine and crystals isomorphous to those of native Efb-C were reproduced using a seleno-l-methionine-labeled form of each mutant protein. Multiwavelength anomalous diffraction (MAD) data were collected on both mutants and analyzed for their phasing power toward solution and refinement of a high-resolution Efb-C crystal structure.
- OSTI ID:
- 22356285
- Journal Information:
- Acta Crystallographica. Section F, Vol. 62, Issue Pt 3; Other Information: PMCID: PMC2197172; PMID: 16511324; PUBLISHER-ID: en5155; OAI: oai:pubmedcentral.nih.gov:2197172; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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