A complete NADPH-cytochrome P450 reductase (CPR) cDNA was isolated fromAnopheles minimus Theobald mosquitoes by using reverse transcription-polymerase chain reaction-based methods. The complete cDNA contains 2,040 bp encoding the protein of 679 amino acids, with a calculated molecular mass of 77.3 kDa. The deduced amino acid sequence had a typical feature of CPR by possessing conserved domains involved in binding of flavin mononucleotide, flavin adenine dinucleotide, and NADPH cofactors. The complete CPR cDNA was expressed as 6x His-tagged fusion protein in both membrane and cytosolic fractions inEscherichia coli, both fractions contained NADPH-cytochromec reducing activity. The membrane-bound form containing N-terminal membrane anchor was subjected to purification, andKm values were determined for NADPH and cytochromec. The purified CPR enzyme was functionally active, as demonstrated by its ability to support CYP6AA3-mediated metabolism in the reconstituted reaction in vitro. Initial test suggested that CYP6AA3 could play a role in deltamethrin metabolism.
How to translate text using browser tools
1 June 2007
Functional Expression of Mosquito NADPH-Cytochrome P450 Reductase inEscherichia coli
Dolnapa Kaewpa,
Soamrutai Boonsuepsakul,
Pornpimol Rongnoparut
ACCESS THE FULL ARTICLE
It is not available for individual sale.
This article is only available to subscribers.
It is not available for individual sale.
It is not available for individual sale.
Journal of Economic Entomology
Vol. 100 • No. 3
June 2007
Vol. 100 • No. 3
June 2007
Anopheles minimus
CYP6AA3
deltamethrin
kinetics
NADPH-cytochrome P450 reductase