pHluorin-assisted expression, purification, crystallization and X-ray diffraction data analysis of the C-terminal domain of the HsdR subunit of the Escherichia coli type I restriction-modification system EcoR124I.
Acta Crystallogr F Struct Biol Commun
; 72(Pt 9): 672-6, 2016 09.
Article
en En
| MEDLINE
| ID: mdl-27599856
The HsdR subunit of the type I restriction-modification system EcoR124I is responsible for the translocation as well as the restriction activity of the whole complex consisting of the HsdR, HsdM and HsdS subunits, and while crystal structures are available for the wild type and several mutants, the C-terminal domain comprising approximately 150 residues was not resolved in any of these structures. Here, three fusion constructs with the GFP variant pHluorin developed to overexpress, purify and crystallize the C-terminal domain of HsdR are reported. The shortest of the three encompassed HsdR residues 887-1038 and yielded crystals that belonged to the orthorhombic space group C2221, with unit-cell parameters a = 83.42, b = 176.58, c = 126.03â
Å, α = ß = γ = 90.00° and two molecules in the asymmetric unit (VM = 2.55â
Å(3)â
Da(-1), solvent content 50.47%). X-ray diffraction data were collected to a resolution of 2.45â
Å.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Proteínas Recombinantes de Fusión
/
Desoxirribonucleasas de Localización Especificada Tipo I
/
Subunidades de Proteína
/
Proteínas de Escherichia coli
/
Proteínas Fluorescentes Verdes
/
Escherichia coli
Idioma:
En
Revista:
Acta Crystallogr F Struct Biol Commun
Año:
2016
Tipo del documento:
Article
País de afiliación:
República Checa
Pais de publicación:
Estados Unidos