Crystallization and preliminary X-ray crystallographic analysis of bacterial tRNA(Sec) in complex with seryl-tRNA synthetase.
Acta Crystallogr Sect F Struct Biol Cryst Commun
; 68(Pt 6): 678-82, 2012 Jun 01.
Article
em En
| MEDLINE
| ID: mdl-22684069
ABSTRACT
Selenocysteine (Sec) is translationally incorporated into proteins in response to the UGA codon. The tRNA specific to Sec (tRNA(Sec)) is first ligated with serine by seryl-tRNA synthetase (SerRS). To elucidate the tertiary structure of bacterial tRNA(Sec) and its specific interaction with SerRS, the bacterial tRNA(Sec) from Aquifex aeolicus was crystallized as the heterologous complex with the archaeal SerRS from Methanopyrus kandleri. Although X-ray diffraction by crystals of tRNA(Sec) in complex with wild-type SerRS was rather poor (to 5.7 Å resolution), the resolution was improved by introducing point mutations targeting the crystal-packing interface. Heavy-atom labelling also contributed to resolution improvement. A 3.2 Å resolution diffraction data set for phase determination was obtained from a K(2)Pt(CN)(4)-soaked crystal.
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Base de dados:
MEDLINE
Assunto principal:
Serina-tRNA Ligase
/
Bactérias
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RNA de Transferência
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Euryarchaeota
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article