Cloning, crystallization and preliminary X-ray diffraction analysis of an intact DNA methyltransferase of a type I restriction-modification enzyme from Vibrio vulnificus.
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, 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%.
Department of Chemistry, Chonnam National University, Gwangju 500-757, Republic of Korea.
Ly
L
Huynh Thi Yen
Department of Chemistry, Chonnam National University, Gwangju 500-757, Republic of Korea.
Suk-Youl
SY
Park
Department of Chemistry, Chonnam National University, Gwangju 500-757, Republic of Korea.
Jeong-Sun
JS
Kim
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods*; D004247:DNA / Q000378:metabolism; D015253:Deoxyribonucleases, Type I Site-Specific / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008745:Methylation; D008969:Molecular Sequence Data; D021122:Protein Subunits; D017386:Sequence Homology, Amino Acid; D017356:Synchrotrons; D041261:Vibrio vulnificus / Q000201:enzymology*
D021122:Protein Subunits; D004247:DNA; D015253:Deoxyribonucleases, Type I Site-Specific
DNA methyltransferases; methylation subunit; specificity subunit; type I restriction and modification