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Crystallization and preliminary X-ray crystallographic analysis of Aquifex aeolicus SelA, a bacterial selenocysteine synthase.
- Source :
-
Acta crystallographica. Section F, Structural biology and crystallization communications [Acta Crystallogr Sect F Struct Biol Cryst Commun] 2012 Sep 01; Vol. 68 (Pt 9), pp. 1128-33. Date of Electronic Publication: 2012 Aug 31. - Publication Year :
- 2012
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Abstract
- Selenocysteine (Sec), the 21st amino acid, is synthesized on its specific tRNA (tRNA(Sec)) via a multi-step process. In bacteria, tRNA(Sec) is ligated first with serine by seryl-tRNA synthetase, which is followed by Ser-to-Sec conversion by Sec synthase (SelA). To elucidate its structure and catalytic mechanism, Aquifex aeolicus SelA was crystallized. Although wild-type SelA crystals diffracted X-rays poorly (to up to 8 Å resolution), the resolution was improved by introducing a quadruple point mutation targeting the loop regions and by methylating the lysine residues, which yielded 3.9 Å resolution diffraction data from a full-length SelA crystal. Truncation of the N-terminal region (ΔN) also improved the resolution. A 3.3 Å resolution data set for phase determination was obtained from a crystal of selenomethionine-substituted Lys-methylated SelA-ΔN.
Details
- Language :
- English
- ISSN :
- 1744-3091
- Volume :
- 68
- Issue :
- Pt 9
- Database :
- MEDLINE
- Journal :
- Acta crystallographica. Section F, Structural biology and crystallization communications
- Publication Type :
- Academic Journal
- Accession number :
- 22949212
- Full Text :
- https://doi.org/10.1107/S1744309112033519