Crystallization and initial X-ray analysis of phenoxazinone synthase from Streptomyces antibioticus

Alex W. Smith, Ana Camara-Artigas, Charles Olea, Wilson A. Francisco, James Paul Allen

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Phenoxazinone synthase, an oligomeric multicopper oxidase produced by Streptomyces antibioticus, is responsible for the six-electron oxidative coupling of two molecules of 4-methyl 3-hydroxy-anthraniloyl pentapeptide to form the phenoxazinone chromophore of the antineoplastic agent actinomycin D. Spectroscopic studies have shown that the enzyme contains one type I (blue) and three to four type II copper centers. However, the exact arrangement of the copper centers in this multicopper oxidase is unknown. As a first step towards determining the three-dimensional structure of the enzyme, phenoxazinone synthase has been crystallized. The hexameric form of phenoxazinone synthase was purified from 72 h cultures of S. lividans containing the plasmid pIJ702. Purified hexamers were concentrated to 75 mg ml-1 and used to grow two forms of crystals. Data collected from the two crystal forms were processed in two separate space groups. Crystals of both forms were grown at 288 K using the sitting-drop vapour-diffusion method. Native data sets extending to resolutions of 3.35 and 2.30 Å have been collected and processed in space groups R32 and P1, respectively.

Original languageEnglish
Pages (from-to)1453-1455
Number of pages3
JournalActa Crystallographica Section D: Biological Crystallography
Volume60
Issue number8
DOIs
Publication statusPublished - Aug 2004

Fingerprint

Streptomyces antibioticus
X ray analysis
Crystallization
oxidase
X-Rays
crystallization
enzymes
actinomycin
Crystals
crystals
Copper
copper
Oxidoreductases
plasmids
x rays
Oxidative Coupling
chromophores
Dactinomycin
Enzymes
Chromophores

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics
  • Condensed Matter Physics
  • Structural Biology

Cite this

Crystallization and initial X-ray analysis of phenoxazinone synthase from Streptomyces antibioticus. / Smith, Alex W.; Camara-Artigas, Ana; Olea, Charles; Francisco, Wilson A.; Allen, James Paul.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 60, No. 8, 08.2004, p. 1453-1455.

Research output: Contribution to journalArticle

Smith, Alex W. ; Camara-Artigas, Ana ; Olea, Charles ; Francisco, Wilson A. ; Allen, James Paul. / Crystallization and initial X-ray analysis of phenoxazinone synthase from Streptomyces antibioticus. In: Acta Crystallographica Section D: Biological Crystallography. 2004 ; Vol. 60, No. 8. pp. 1453-1455.
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