Purification, crystallization and preliminary crystallographic analysis of DR0248, an MNT-HEPN fused protein from Deinococcus radiodurans.
Acta Crystallogr F Struct Biol Commun
; 71(Pt 1): 49-53, 2015 Jan 01.
Article
em En
| MEDLINE
| ID: mdl-25615968
ABSTRACT
DR0248 is a protein identified in the Deinococcus radiodurans (DR) genome that is predicted to encompass two domains an N-terminal minimal nucleotidyl transferase domain (MNT) and a C-terminal higher eukaryotes and prokaryotes nucleotide-binding domain (HEPN). These two domains, usually encoded in two ORFs, have been suggested to play the role of a toxin-antitoxin (TA) system in prokaryotes. Recombinant DR0248 was overexpressed and purified from Escherichia coli and diffraction-quality crystals were obtained in the presence of the detergent molecules dodecyldimethylamine oxide (DDAO) and octaethylene glycol monododecyl ether (C12E8), which were used as crystallization additives. Crystals grown with DDAO diffracted to a resolution of 2.24â
Å and belonged to space group C222(1), with unit-cell parameters a=98.4, b=129.9, c=59.2â
Å. Crystals grown with C12E8 diffracted to a resolution of 1.83â
Å and belonged to space group P2(1)2(1)2(1), with unit-cell parameters a=51.6, b=87.2, c=108.2â
Å. The structure was solved by multiwavelength anomalous dispersion from zinc bound to the protein using a single crystal obtained in the presence of DDAO.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
/
Deinococcus
/
Nucleotidiltransferases
Idioma:
En
Revista:
Acta Crystallogr F Struct Biol Commun
Ano de publicação:
2015
Tipo de documento:
Article
País de afiliação:
França