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Cloning, crystallization and preliminary X‐ray diffraction analysis of an intact DNA methyltransferase of a type I restriction–modification enzyme from Vibrio vulnificus
Author(s) -
Huynh Thi Yen Ly,
Park SukYoul,
Kim JeongSun
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14004543
Subject(s) - crystallography , crystallization , monoclinic crystal system , polyethylene glycol , methylation , ammonium sulfate , crystal (programming language) , methyltransferase , chemistry , biology , materials science , crystal structure , dna , biochemistry , organic chemistry , computer science , programming language
Independently of the restriction (HsdR) subunit, the specificity (HsdS) and methylation (HsdM) subunits interact with each other, and function as a methyltransferase in type I restriction–modification systems. A single gene that combines the HsdS and HsdM subunits in Vibrio vulnificus YJ016 was expressed and purified. A crystal suitable for X‐ray diffraction was obtained from 25%( w / v ) polyethylene glycol monomethylether 5000, 0.1 M HEPES pH 8.0, 0.2 M ammonium sulfate at 291 K by hanging‐drop vapour diffusion. Diffraction data were collected to a resolution of 2.31 Å using synchrotron radiation. The crystal belonged to the primitive monoclinic space group P 2 1 , with unit‐cell parameters a = 93.25, b = 133.04, c = 121.49 Å, β = 109.7°. With four molecules in the asymmetric unit, the crystal volume per unit protein weight was 2.61 Å 3 Da −1 , corresponding to a solvent content of 53%.