Crystallization and preliminary x-ray diffraction analysis of a recombinant bacterial heme oxygenase (Hmu O) from Corynebacterium diphtheriae

Grace C. Chu, Sam Yong Park, Yoshitsugu Shiro, Tadashi Yoshida, Masao Ikeda-Saito

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)

Abstract

Hmu O is a 24-kDa soluble bacterial heme degradation enzyme found in the pathogen Corynebaeterium diphtheriae, the causative agent of diphtheria. Similar to the mammalian heme oxygenase, it binds hemin stoichiometrically and catalyzes the oxygen-dependent conversion of hemin to biliverdin, carbon monoxide, and free iron. Iron is an essential nutrient for bacteria and especially important for pathogenesis. Here we report the first crystallization and preliminary crystallographic study of the heme-Hmu O complex formed from hemin and a recombinant Hmu O, which was expressed in Escherichia coil from a synthetic gene based on the putative hmu O gene sequence. Crystals of the heme-Hmu O complex were obtained by the sitting drop vapor diffusion method using a precipitant solution containing 18% (w/v) PEG 8000 and 0.2 M calcium acetate in 0.1 M sodium cacodylate (pH 6.5). Using synchrotron radiation, the heme-Hmu O crystal diffracted to 2.8 Å resolution. It belongs to the monoclinic space group C2, with unit cell parameters a = 123.18 Å, b = 44.51 Å, c = 92.10 Å, and β = 123.3°. Assuming one molecule of the heme-Hmu O complex per asymmetric unit, the calculated value of V(m) is 2.89 Å3/Da.

Original languageEnglish
Pages (from-to)171-174
Number of pages4
JournalJournal of Structural Biology
Volume126
Issue number2
DOIs
Publication statusPublished - 1999 Jun 15

Keywords

  • Crystallization
  • Heme oxygenase
  • Hmu O
  • X-ray diffraction

ASJC Scopus subject areas

  • Structural Biology

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