Resonance Raman evidence for a novel charge relay activation mechanism of the CO-dependent heme protein transcription factor CooA

Kathleen M. Vogel, Thomas G. Spiro, Daniel Shelver, Marc V. Thorsteinsson, Gary P. Roberts

Research output: Contribution to journalArticlepeer-review

71 Scopus citations

Abstract

Resonance Raman spectra of the CO-responsive transcription factor CooA from Rhodospirillum rubrum provides evidence on the nature of heme ligation and its CO activation mechanism. The Fe(III) form gives standard low-spin heme spectrum, while the Fe(II) form is low spin for wild-type (WT) CooA and mixed spin for a CooA variant, H77Y, with an His77Tyr substitution. The Fe(II) porphyrin skeletal mode v11 is at a value (1541 cm-1) indicative of a neutral donor ligand for the H77Y variant but is at an unusually depressed frequency (1529 cm-1) for the WT protein, indicating a strongly donating ligand. This ligand is proposed to be His77 imidazolate, formed by proton transfer to a nearby acceptor. The WT CO adduct has FeCO and CO stretching frequencies that indicate a neutral trans ligand and negative polarity in the CO binding pocket, while the CO adduct of His77Tyr has both 6- and 5-coordinate signals and a nonpolar CO environment. Photolysis of the WT CO adduct by the Raman laser produced a low-spin product at steady state, indicating fast recombination of the displaced ligand. These data suggest a novel YH - His- charge relay mechanism for CooA activation by CO. In this mechanism, His77 is reprotonated upon ligand displacement by CO; CO displaces either His77 or the trans ligand, X. The resulting charge on Y- may induce the protein conformation change required for site-selective DNA binding.

Original languageEnglish (US)
Pages (from-to)2679-2687
Number of pages9
JournalBiochemistry
Volume38
Issue number9
DOIs
StatePublished - Mar 2 1999
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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