CO-dependent activity-controlling mechanism of heme-containing CO-sensor protein, neuronal PAS domain protein 2

Takeshi Uchida, Emiko Sato, Akira Sato, Ikuko Sagami, Toru Shimizu, Teizo Kitagawa

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68 Citations (Scopus)


Neuronal PAS domain protein 2, which was recently established to be a heme protein, acts as a CO-dependent transcription factor. The protein consists of the basic helix-loop-helix domain and two heme-containing PAS domains (PAS-A and PAS-B). In this study, we prepared wild type and mutants of the isolated PAS-A domain and measured resonance Raman spectra of these proteins. Upon excitation of the Raman spectrum at 363.8 nm, a band assignable to Fe3+-S stretching was observed at 334 cm-1 for the ferric wild type protein; in contrast, this band was drastically weaker in the spectrum of C170A, suggesting that Cys170 is an axial ligand of the ferric heme. The Raman spectrum of the reduced form of wild type was mainly of six-coordinate low spin, and the ν11 band, which is sensitive to the donor strength of the axial ligand, was lower than that of reduced cytochrome c3, suggesting coordination of a strong ligand and thus a deprotonated His. In the reduced forms of H119A and H171A, the five-coordinate species became more prevalent, whereas no such changes were observed for C170A, indicating that His119 and His171, but not Cys170, are axial ligands in the ferrous heme. This means that ligand replacement from Cys to His occurs upon heme reduction. The νFe-CO versus νC-O correlation indicates that a neutral His is a trans ligand of CO. Our results support a mechanism in which CO binding disrupts the hydrogen bonding of His171 with surrounding amino acids, which induces conformational changes in the His171-Cys170 moiety, leading to physiological signaling.

Original languageEnglish
Pages (from-to)21358-21368
Number of pages11
JournalJournal of Biological Chemistry
Issue number22
Publication statusPublished - 2005 Jun 3

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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