Crystallization of recombinant bifunctional nuclease TBN1 from tomato
Language English Country England, Great Britain Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
21206042
PubMed Central
PMC3079990
DOI
10.1107/s1744309110048177
PII: S1744309110048177
Knihovny.cz E-resources
- MeSH
- Deoxyribonucleases chemistry genetics MeSH
- Ions chemistry MeSH
- Protein Conformation MeSH
- Crystallization MeSH
- Crystallography, X-Ray MeSH
- Molecular Sequence Data MeSH
- Recombinant Proteins chemistry genetics MeSH
- Plant Proteins chemistry genetics MeSH
- Solanum lycopersicum chemistry genetics MeSH
- Zinc chemistry MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Deoxyribonucleases MeSH
- Ions MeSH
- Recombinant Proteins MeSH
- Plant Proteins MeSH
- Zinc MeSH
The endonuclease TBN1 from Solanum lycopersicum (tomato) was expressed in Nicotiana benthamiana leaves and purified with suitable quality and in suitable quantities for crystallization experiments. Two crystal forms (orthorhombic and rhombohedral) were obtained and X-ray diffraction experiments were performed. The presence of natively bound Zn2+ ions was confirmed by X-ray fluorescence and by an absorption-edge scan. X-ray diffraction data were collected from the orthorhombic (resolution of 5.2 Å) and rhombohedral (best resolution of 3.2 Å) crystal forms. SAD, MAD and MR methods were applied for solution of the phase problem, with partial success. TBN1 contains three Zn2+ ions in a similar spatial arrangement to that observed in nuclease P1 from Penicillium citrinum.
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