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.
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Acknowledgment
This work was supported by National Institutes of Health Grant GM25515.
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Straub, W.E., Nishida, C.R., de Montellano, P.R.O. (2013). Expression in Escherichia coli of a Cytochrome P450 Enzyme with a Cobalt Protoporphyrin IX Prosthetic Group. In: Phillips, I., Shephard, E., Ortiz de Montellano, P. (eds) Cytochrome P450 Protocols. Methods in Molecular Biology, vol 987. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-321-3_9
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DOI: https://doi.org/10.1007/978-1-62703-321-3_9
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-320-6
Online ISBN: 978-1-62703-321-3
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