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

Supangat Supangat, Young Jun An, Younguk Sun, Suk Tae Kwon, Sun Shin Cha

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

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 l 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, Β = 82.4, = 74.6°. Assuming the presence of two molecules in the unit cell, the solvent content was estimated to be about 53.4%.

Original languageEnglish
Pages (from-to)1583-1585
Number of pages3
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume66
Issue number12
DOIs
StatePublished - Dec 2010

Keywords

  • DNA ligases
  • multiple cofactors
  • Sulfophobococcus zilligii

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