Histidine 20, the crucial proximal axial heme ligand of bacterial heme oxygenase Hmu O from Corynebacterium diphtheriae

Grace C. Chu, Koki Katakura, Takeshi Tomita, Xuhong Zhang, Danyu Sun, Michihiko Sato, Masanao Sasahara, Takamasa Kayama, Masao Ikeda-Saito, Tadashi Yoshida

研究成果: Article査読

28 被引用数 (Scopus)

抄録

The hemin complex of Hmu O, a 24-kDa soluble heme degradation enzyme in Corynebacterium diphtheriae, is coordinated axially to a neutral imidazole of a proximal histidine residue in Hmu O. To identify which of the eight histidines in Hmu O is the proximal heme ligand, we have constructed and expressed the plasmids for eight His → Ala Hmu O mutants. Reconstituted with hemin, the active site structures and enzymatic activity of these mutants have been examined by EPR, resonance Raman, and optical absorption spectroscopy. EPR of the NO-bound ferrous heme-Hmu O mutant complexes reveals His20 as the proximal heme ligand in Hmu O, and this is confirmed by resonance Raman results from the ligand-free ferrous heme-H20A. All eight His → Ala mutants bind hemin stoichiometrically, proving that none of the histidines is essential for hemin-Hmu O formation. However, His20 is crucial to Hmu O catalysis. Its absence by point mutation has inhibited the conversion of hemin to biliverdin. The ferric heme-H20A complex is pentacoordinate. Resonance Raman of the CO-bound ferrous heme-H20A corroborates this and reveals an Fe-C-O bending mode, δ(Fe-C-O), the first reported for a pentacoordinate CO-bound hemeprotein. The appearance of δ(Fe- C-O) in C. diphtheriae Hmu O H20A but not mammalian HO-1 mutant H25A indicates that the heme environment between the two heme oxygenases is different.

本文言語English
ページ(範囲)17494-17500
ページ数7
ジャーナルJournal of Biological Chemistry
275
23
DOI
出版ステータスPublished - 2000 6月 9

ASJC Scopus subject areas

  • 生化学
  • 分子生物学
  • 細胞生物学

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