Isotopic labeling of the heme cofactor in cytochrome p450 and other heme proteins.

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
David Irby Bryson (Creator)
Sandhya C. Manjunath (Creator)
Gregory M. Raner, Associate Professor and Graduate Director (Creator)
The University of North Carolina at Greensboro (UNCG )
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Abstract: A recombinant bacterial expression system that generates 13C-labeled heme or 15N-labeled heme in functional cytochrome P450 enzymes and other heme-containing systems is reported here using a mutant strain of Escherichia coli (HU227) in which the HemA gene is inactive. By synthesizing several isotopomers of aminolevulinic acid with 13C or 15N at different locations, isotopes have been incorporated with high abundance into the heme cofactor of five different cytochrome P450 isoforms, along with one peroxidase. Confirmed both 13C- and 15N-incorporation; spectral and catalytic assays show the labeled enzymes produced in this system are functional.

Additional Information

Language: English
Date: 2011
aminolevulinic acid, Carbon-13, Cytochrome P450, Heme, Isotopic labeling, Nitrogen-15, biochemistry, biotechnology

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