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Purification, crystallization and preliminary crystallographic analysis of a multiple cofactor-dependent DNA ligase from Sulfophobococcus zilligii.

ABSTRACT: A recombinant DNA ligase from Sulfophobococcus zilligii that shows multiple cofactor specificity (ATP, ADP and GTP) was expressed in Escherichia coli and purified under reducing conditions. Crystals were obtained by the microbatch crystallization method at 295?K in a drop containing 1?µl protein solution (10?mg?ml(-1)) and an equal volume of mother liquor [0.1?M HEPES pH 7.5, 10%(w/v) polyethylene glycol 10?000]. A data set was collected to 2.9?Å resolution using synchrotron radiation. The crystals belonged to space group P1, with unit-cell parameters a=63.7, b=77.1, c=77.8?Å, ?=83.4, ?=82.4, ?=74.6°. Assuming the presence of two molecules in the unit cell, the solvent content was estimated to be about 53.4%.

SUBMITTER: Supangat S

PROVIDER: S-EPMC2998359 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature

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Purification, crystallization and preliminary crystallographic analysis of a multiple cofactor-dependent DNA ligase from Sulfophobococcus zilligii.

Supangat Supangat S An Young Jun YJ Sun Younguk Y Kwon Suk-Tae ST Cha Sun-Shin SS

Acta crystallographica. Section F, Structural biology and crystallization communications 20101116 Pt 12

A recombinant DNA ligase from Sulfophobococcus zilligii that shows multiple cofactor specificity (ATP, ADP and GTP) was expressed in Escherichia coli and purified under reducing conditions. Crystals were obtained by the microbatch crystallization method at 295 K in a drop containing 1 µl protein solution (10 mg ml(-1)) and an equal volume of mother liquor [0.1 M HEPES pH 7.5, 10%(w/v) polyethylene glycol 10 000]. A data set was collected to 2.9 Å resolution using synchrotron radiation. The cryst ...[more]

PMID: 21139200

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