Combining site-specific mutagenesis and seeding as a strategy to crystallize ‘difficult’ proteins: the case of Staphylococcus aureus thioredoxin
Journal Article
·
· Acta Crystallographica. Section F
- Brussels Center for Redox Biology, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB) at the Vrije Universiteit Brussel (VUB), Pleinlaan 2, 1050 Brussel (Belgium)
- Architecture et Fonction des Macromolecules Biologiques, UMR 6098 CNRS and Universites d’Aix-Marseille I et II, Campus de Luminy, 163 Avenue de Luminy 13288 Marseille CEDEX 9 (France)
- Department of Molecular and Cellular Interactions, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB) at the Vrije Universiteit Brussel (VUB), Pleinlaan 2, 1050 Brussel (Belgium)
S. aureus thioredoxin was crystallized using a combination of seeding and site-specific mutagenesis. The P31T mutant of Staphylococcus aureus thioredoxin crystallizes spontaneously in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 41.7, b = 49.5, c = 55.6 Å. The crystals diffract to 2.2 Å resolution. Isomorphous crystals of wild-type thioredoxin as well as of other point mutants only grow when seeded with the P31T mutant. These results suggest seeding as a valuable tool complementing surface engineering for proteins that are hard to crystallize.
- OSTI ID:
- 22356421
- Journal Information:
- Acta Crystallographica. Section F, Vol. 62, Issue Pt 12; Other Information: PMCID: PMC2225371; PMID: 17142910; PUBLISHER-ID: ll5093; OAI: oai:pubmedcentral.nih.gov:2225371; 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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