Evidence for basic ferryls in cytochromes P450

Rachel K. Behan, Lee M. Hoffart, Kari L. Stone, Carsten Krebs, Michael T. Green

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

Using a combination of Mössbauer spectroscopy and density functional calculations, we have determined that the ferryl forms of P450BM3 and P450cam are protonated at physiological pH. Density functional calculations were performed on large active-site models of these enzymes to determine the theoretical Mössbauer parameters for the ferryl and protonated ferryl (FeIVOH) species. These calculations revealed a significant enlargement of the quadrupole splitting parameter upon protonation of the ferryl unit. The calculated quadrupole splittings for the protonated and unprotonated ferryl forms of P450BM3 are ΔEQ = 2.17 mm/s and ΔEQ = 1.05mm/s, respectively. For P450cam, they are ΔEQ = 1.84 mm/s and ΔEQ = 0.66 mm/s, respectively. The experimentally determined quadrupole splittings (P450 BM3, ΔEQ = 2.16 mm/s; P450cam, ΔEQ = 2.06 mm/s) are in good agreement with the values calculated for the protonated forms of the enzymes. Our results suggest that basic ferryls are a natural consequence of thiolate-ligated hemes.

Original languageEnglish (US)
Pages (from-to)11471-11474
Number of pages4
JournalJournal of the American Chemical Society
Volume128
Issue number35
DOIs
StatePublished - Sep 6 2006

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Camphor 5-Monooxygenase
Cytochrome P-450 Enzyme System
Density functional theory
Enzymes
Protonation
Spectroscopy
Heme
Catalytic Domain
Spectrum Analysis

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Behan, R. K., Hoffart, L. M., Stone, K. L., Krebs, C., & Green, M. T. (2006). Evidence for basic ferryls in cytochromes P450. Journal of the American Chemical Society, 128(35), 11471-11474. https://doi.org/10.1021/ja062428p
Behan, Rachel K. ; Hoffart, Lee M. ; Stone, Kari L. ; Krebs, Carsten ; Green, Michael T. / Evidence for basic ferryls in cytochromes P450. In: Journal of the American Chemical Society. 2006 ; Vol. 128, No. 35. pp. 11471-11474.
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Behan, RK, Hoffart, LM, Stone, KL, Krebs, C & Green, MT 2006, 'Evidence for basic ferryls in cytochromes P450', Journal of the American Chemical Society, vol. 128, no. 35, pp. 11471-11474. https://doi.org/10.1021/ja062428p

Evidence for basic ferryls in cytochromes P450. / Behan, Rachel K.; Hoffart, Lee M.; Stone, Kari L.; Krebs, Carsten; Green, Michael T.

In: Journal of the American Chemical Society, Vol. 128, No. 35, 06.09.2006, p. 11471-11474.

Research output: Contribution to journalArticle

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N2 - Using a combination of Mössbauer spectroscopy and density functional calculations, we have determined that the ferryl forms of P450BM3 and P450cam are protonated at physiological pH. Density functional calculations were performed on large active-site models of these enzymes to determine the theoretical Mössbauer parameters for the ferryl and protonated ferryl (FeIVOH) species. These calculations revealed a significant enlargement of the quadrupole splitting parameter upon protonation of the ferryl unit. The calculated quadrupole splittings for the protonated and unprotonated ferryl forms of P450BM3 are ΔEQ = 2.17 mm/s and ΔEQ = 1.05mm/s, respectively. For P450cam, they are ΔEQ = 1.84 mm/s and ΔEQ = 0.66 mm/s, respectively. The experimentally determined quadrupole splittings (P450 BM3, ΔEQ = 2.16 mm/s; P450cam, ΔEQ = 2.06 mm/s) are in good agreement with the values calculated for the protonated forms of the enzymes. Our results suggest that basic ferryls are a natural consequence of thiolate-ligated hemes.

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