Protocol

Cytochrome P450 Protocols

Volume 987 of the series Methods in Molecular Biology pp 107-113

Date:

Expression in Escherichia coli of a Cytochrome P450 Enzyme with a Cobalt Protoporphyrin IX Prosthetic Group

  • Wesley E. StraubAffiliated withLife Technologies
  • , Clinton R. NishidaAffiliated withDepartment of Pharmaceutical Chemistry, University of California
  • , Paul R. Ortiz de MontellanoAffiliated withDepartment of Pharmaceutical Chemistry, University of California

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Abstract

Unlike many hemoproteins, the prosthetic heme group of most cytochrome P450 enzymes cannot be extracted and replaced by modified heme groups. Here, we describe a procedure for generating a cytochrome P450 enzyme (CYP119) with cobalt protoporphyrin IX as its prosthetic group. This is achieved by expressing the protein in Escherichia coli in iron-limited medium and adding cobalt to the medium at the moment that inducible protein expression is initiated.

Key words

Cobalt protoporphyrin IX Cobalt heme CYP119 Heme modification Bacterial ferrochelatase