Crystallization and diffraction data of 1H-3-hydroxy-4-oxoquinoline 2,4-dioxygenase: a cofactor-free oxygenase of the alpha/beta-hydrolase family

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 May 1;63(Pt 5):378-81. doi: 10.1107/S1744309107013760. Epub 2007 Apr 6.

Abstract

1H-3-Hydroxy-4-oxoquinoline 2,4-dioxygenase (QDO) from Pseudomonas putida 33/1 catalyses the oxygenolysis of 1H-3-hydroxy-4-oxoquinoline to form N-formylanthranilic acid and carbon monoxide without the aid of cofactors. Both N-terminally His6-tagged and native QDO were overexpressed in Escherichia coli and purified by conventional chromatographic procedures. Untagged QDO, but not His6-tagged QDO, was crystallized by the vapour-diffusion method, giving hexagonal bipyramid crystals belonging to space group P6(1)22. Selenomethionine-containing native QDO was prepared and crystallized under identical conditions. The unit-cell parameters were a = b = 90.1, c = 168.6 A, alpha = beta = 90, gamma = 120 degrees. Using synchrotron radiation, these crystals diffract to 2.5 A. The expression, purification and crystallization of QDO are reported here.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Arthrobacter / enzymology
  • Base Sequence
  • Crystallization
  • Crystallography, X-Ray
  • DNA Primers
  • Dioxygenases / chemistry*
  • Dioxygenases / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Hydrolases / chemistry*
  • Protein Conformation
  • Pseudomonas putida / enzymology
  • Substrate Specificity

Substances

  • DNA Primers
  • 1H-3-hydroxy-4-oxoquinaldine 2,4-dioxygenase (decyclizing, CO-forming)
  • Dioxygenases
  • Hydrolases