Cleaved thioredoxin fusion protein enables the crystallization of poorly soluble ERα in complex with synthetic ligands
- Laboratoire de Biologie et Génomique Structurales, IGBMC, Illkirch (France)
- Department of Biochemistry, University of Cambridge (United Kingdom)
A new crystallization strategy: the presence of cleaved thioredoxin fusion is critical for crystallization of the estrogen nuclear receptor ligand binding domain in complex with synthetic ligands. This novel technique should be regarded as an interesting alternative for crystallization of difficult proteins. The ligand-binding domain (LBD) of human oestrogen receptor α was produced in Escherichia coli as a cleavable thioredoxin (Trx) fusion in order to improve solubility. Crystallization trials with either cleaved and purified LBD or with the purified fusion protein both failed to produce crystals. In another attempt, Trx was not removed from the LBD after endoproteolytic cleavage and its presence promoted nucleation and subsequent crystal growth, which allowed the structure determination of two different LBD–ligand–coactivator peptide complexes at 2.3 Å resolution. This technique is likely to be applicable to other low-solubility proteins.
- OSTI ID:
- 22360461
- Journal Information:
- Acta Crystallographica. Section F, Vol. 64, Issue Pt 1; Other Information: PMCID: PMC2373989; PMID: 18097104; PUBLISHER-ID: bw5218; OAI: oai:pubmedcentral.nih.gov:2373989; 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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