Heme-based sensing by the mammalian circadian protein CLOCK

Gudrun S. Lukat-Rodgers, Cristina Correia, Maria Victoria Botuyan, Georges Mer, Kenton R. Rodgers

Research output: Contribution to journalArticle

39 Scopus citations

Abstract

Heme is emerging as a key player in the synchrony of circadian-coupled transcriptional regulation. Current evidence suggests that levels of circadian-linked transcription are regulated in response to both the availability of intracellular heme and heme-based sensing of carbon monoxide (CO) and possibly nitric oxide (NO). The protein CLOCK is central to the regulation and maintenance of circadian rhythms in mammals. CLOCK comprises two PAS domains, each with a heme binding site. Our studies focus on the functionality of the murine CLOCK PAS-A domain (residues 103-265). We show that CLOCK PAS-A binds iron(III) protoporhyrin IX to form a complex with 1:1 stoichiometry. Optical absorbance and resonance Raman studies reveal that the heme of ferric CLOCK PAS-A is a six-coordinate, low-spin complex whose resonance Raman signature is insensitive to pH over the range of protein stability. Ferrous CLOCK PAS-A is a mixture of five-coordinate, high-spin and six-coordinate, low-spin complexes. Ferrous CLOCK PAS-A forms complexes with CO and NO. Ferric CLOCK PAS-A undergoes reductive nitrosylation in the presence of NO to generate a CLOCK PAS-A-NO, which is a five-coordinate {FeNO} 7 complex. Formation of the highly stable {FeNO} 7 heme complex from either ferrous or ferric heme makes possible the binding of NO at very low concentration, a characteristic of NO sensors. Comparison of the spectroscopic properties and CO-binding kinetics of CLOCK PAS-A with other CO sensor proteins reveals that CLOCK PAS-A exhibits chemical properties consistent with a heme-based gas sensor protein.

Original languageEnglish (US)
Pages (from-to)6349-6365
Number of pages17
JournalInorganic Chemistry
Volume49
Issue number14
DOIs
StatePublished - Jul 19 2010

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Inorganic Chemistry

Fingerprint Dive into the research topics of 'Heme-based sensing by the mammalian circadian protein CLOCK'. Together they form a unique fingerprint.

  • Cite this

    Lukat-Rodgers, G. S., Correia, C., Botuyan, M. V., Mer, G., & Rodgers, K. R. (2010). Heme-based sensing by the mammalian circadian protein CLOCK. Inorganic Chemistry, 49(14), 6349-6365. https://doi.org/10.1021/ic902388q