Acetylenes: cytochrome P450 oxidation and mechanism-based enzyme inactivation

Abstract

The oxidation of carbon–carbon triple bonds by cytochrome P450 produces ketene metabolites that are hydrolyzed to acetic acid derivatives or are trapped by nucleophiles. In the special case of 17α-ethynyl sterols, D-ring expansion and de-ethynylation have been observed as competing pathways. The oxidation of acetylenic groups is also associated with mechanism-based inactivation of cytochrome P450 enzymes. One mechanism for this inactivation is reaction of the ketene metabolite with cytochrome P450 residues essential for substrate binding or catalysis. However, in the case of monosubstituted acetylenes, inactivation can also occur by addition of the oxidized acetylenic function to a nitrogen of the heme prosthetic group. This addition reaction is not mediated by the ketene metabolite, but rather occurs during oxygen transfer to the triple bond. In some instances, a detectable intermediate is formed that is most consistent with a ketocarbene–iron heme complex. This complex can progress to the N-alkylated heme or revert back to the unmodified enzyme. The ketocarbene complex may intervene in the formation of all the N-alkyl heme adducts, but is normally too unstable to be detected.

Keywords:

• Acetylene oxidation
• ketene formation
• cytochrome P450 inactivation
• iron–carbene complexes
• oxirene
• ethynylsterols

Disclosure statement

No potential conflict of interest was reported by the author.

Additional information

Funding

This work was financially supported by Grant AI074824 from the National Institutes of Health.