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Expression, crystallization and preliminary X-ray diffraction analysis of a modification subunit of a putative type I restriction enzyme from Vibrio vulnificus YJ016.

ABSTRACT: 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%.

SUBMITTER: Lee HJ

PROVIDER: S-EPMC2802878 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature

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Expression, crystallization and preliminary X-ray diffraction analysis of a modification subunit of a putative type I restriction enzyme from Vibrio vulnificus YJ016.

Lee Hyun Ju HJ Nishi Kosuke K Song Jung Mi JM Kim Jeong Sun JS

Acta crystallographica. Section F, Structural biology and crystallization communications 20091127 Pt 12

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 ...[more]

PMID: 20054126

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