Expression, crystallization and preliminary X-ray diffraction analysis of a modification subunit of a putative type I restriction enzyme from Vibrio vulnificus YJ016.
Modification (HsdM) and specificity (HsdS) subunits are constituents of an active methyltransferase (MTase) of multifunctional type I restriction enzymes. To provide a molecular background on HsdM, a putative hsdM gene from Vibrio vulnificus YJ016 (HsdM_Vv) was cloned and the expressed protein was purified and crystallized from 22%(w/v) polyethylene glycol 8000, 0.02 M imidazole pH 7.5 and 5 mM beta-mercaptoethanol. Diffraction data were collected to 1.86 A resolution using synchrotron radiation. The crystal belonged to the tetragonal space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 78.9, c = 165.8 A. With one molecule in the asymmetric unit, the crystal volume per unit protein weight was 2.12 A(3) Da(-1), with a solvent content of 42%.
Lee
Hyun Ju
HJ
Department of Chemistry, Chonnam National University, Gwangju 500-757, Republic of Korea.
Nishi
Kosuke
K
Song
Jung Mi
JM
Kim
Jeong Sun
JS
D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D015253:Deoxyribonucleases, Type I Site-Specific / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D041261:Vibrio vulnificus / Q000201:enzymology* / Q000235:genetics
D021122:Protein Subunits; D011994:Recombinant Proteins; D015253:Deoxyribonucleases, Type I Site-Specific