Formation of heme-derived products by the reaction of ferrous deoxymyoglobin with BrCCl3

Yoichi Osawa, Robert J. Highet, Constance M. Murphy, Robert J. Cotter, Lance R. Pohl

Research output: Contribution to journalArticle

Abstract

The reductive metabolism of BrCCl3 by ferrous deoxymyoglobin leads to the formation of heme-derived products, including those that are covalently bound to the protein. BrCCl3 caused a decrease in the absorption of the oxidized myoglobin in the Soret region, suggesting that the prosthetic heme moiety had been modified. Examination of the total reaction mixture by HPLC revealed the presence of three major soluble heme-derived products and a protein-bound heme adduct. The soluble products were unequivocally identified by electronic absorption, FAB mass spectrometry, and 2D NOESY and COSY NMR as β-carboxyvinyl, α-hydroxy-β-(trichloromethyl)ethyl, and α,β-bis(trichloromethyl)ethyl derivatives of the heme prosthetic group and are believed to result from the initial regiospecific attack of the trichloromethyl radical at the ring I vinyl substituent. As might be expected from the bulk of the trichloromethyl group and the crowded environment of the heme, the α,β-bis(trichloromethyl)ethyl moiety was held in a rigid conformation about the heme. A similar approach utilizing cytochrome P-450 instead of myoglobin may be used to study the mechanism of inactivation of the P-450 enzyme by the trichloromethyl radical.

Original languageEnglish (US)
Pages (from-to)4462-4467
Number of pages6
JournalJournal of the American Chemical Society
Volume111
Issue number12
StatePublished - 1989

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Prosthetics
Heme
Proteins
Metabolism
Mass spectrometry
Conformations
Nuclear magnetic resonance
Derivatives
Myoglobin
Cytochrome P-450 Enzyme System
deoxymyoglobin
Mass Spectrometry
High Pressure Liquid Chromatography

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Formation of heme-derived products by the reaction of ferrous deoxymyoglobin with BrCCl3. / Osawa, Yoichi; Highet, Robert J.; Murphy, Constance M.; Cotter, Robert J.; Pohl, Lance R.

In: Journal of the American Chemical Society, Vol. 111, No. 12, 1989, p. 4462-4467.

Research output: Contribution to journalArticle

Osawa, Yoichi ; Highet, Robert J. ; Murphy, Constance M. ; Cotter, Robert J. ; Pohl, Lance R. / Formation of heme-derived products by the reaction of ferrous deoxymyoglobin with BrCCl3. In: Journal of the American Chemical Society. 1989 ; Vol. 111, No. 12. pp. 4462-4467.
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AU - Pohl, Lance R.

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