Cloning, crystallization and preliminary X-ray diffraction analysis of an intact DNA methyltransferase of a type I restriction-modification enzyme from Vibrio vulnificus.

ABSTRACT: 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 P21, with unit-cell parameters a = 93.25, b = 133.04, c = 121.49?Å, ? = 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%.

SUBMITTER: Huynh Thi Yen L 

PROVIDER: S-EPMC3976070 | biostudies-literature | 2014 Apr

REPOSITORIES: biostudies-literature