Next Article in Journal
The Combination of Catechin and Epicatechin Gallate from Fructus Crataegi Potentiates β-Lactam Antibiotics Against Methicillin-Resistant Staphylococcus aureus (MRSA) in Vitro and in Vivo
Next Article in Special Issue
MICAL, the Flavoenzyme Participating in Cytoskeleton Dynamics
Previous Article in Journal
Stable Isolation of Phycocyanin from Spirulina platensis Associated with High-Pressure Extraction Process
Previous Article in Special Issue
Crystal Structure of Dimeric Flavodoxin from Desulfovibrio gigas Suggests a Potential Binding Region for the Electron-Transferring Partner
Article Menu

Export Article

Open AccessArticle
Int. J. Mol. Sci. 2013, 14(1), 1788-1801;

Modeling of Anopheles minimus Mosquito NADPH-Cytochrome P450 Oxidoreductase (CYPOR) and Mutagenesis Analysis

Department of Biochemistry, Faculty of Science, Burapha University, Chonburi 20131, Thailand
Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
Faculty of Animal Sciences and Agricultural Technology, Silpakorn University, Petchaburi IT Campus, Petchaburi 76120, Thailand
Author to whom correspondence should be addressed.
Received: 25 September 2012 / Revised: 19 November 2012 / Accepted: 5 January 2013 / Published: 16 January 2013
(This article belongs to the Special Issue Flavins)
Full-Text   |   PDF [979 KB, uploaded 19 June 2014]   |  


Malaria is one of the most dangerous mosquito-borne diseases in many tropical countries, including Thailand. Studies in a deltamethrin resistant strain of Anopheles minimus mosquito, suggest cytochrome P450 enzymes contribute to the detoxification of pyrethroid insecticides. Purified A. minimus CYPOR enzyme (AnCYPOR), which is the redox partner of cytochrome P450s, loses flavin-adenosine di-nucleotide (FAD) and FLAVIN mono-nucleotide (FMN) cofactors that affect its enzyme activity. Replacement of leucine residues at positions 86 and 219 with phenylalanines in FMN binding domain increases FMN binding, enzyme stability, and cytochrome c reduction activity. Membrane-Bound L86F/L219F-AnCYPOR increases A. minimus P450-mediated pyrethroid metabolism in vitro. In this study, we constructed a comparative model structure of AnCYPOR using a rat CYPOR structure as a template. Overall model structure is similar to rat CYPOR, with some prominent differences. Based on primary sequence and structural analysis of rat and A. minimus CYPOR, C427R, W678A, and W678H mutations were generated together with L86F/L219F resulting in three soluble Δ55 triple mutants. The C427R triple AnCYPOR mutant retained a higher amount of FAD binding and increased cytochrome c reduction activity compared to wild-type and L86F/L219F-Δ55AnCYPOR double mutant. However W678A and W678H mutations did not increase FAD and NAD(P)H bindings. The L86F/L219F double and C427R triple membrane-bound AnCYPOR mutants supported benzyloxyresorufin O-deakylation (BROD) mediated by mosquito CYP6AA3 with a two- to three-fold increase in efficiency over wild-type AnCYPOR. The use of rat CYPOR in place of AnCYPOR most efficiently supported CYP6AA3-mediated BROD compared to all AnCYPORs. View Full-Text
Keywords: Anopheles minimus mosquito; cytochrome P450 oxidoreductase (CYPOR); structure; FAD and NAD(P)H bindings Anopheles minimus mosquito; cytochrome P450 oxidoreductase (CYPOR); structure; FAD and NAD(P)H bindings

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

Supplementary material


Share & Cite This Article

MDPI and ACS Style

Sarapusit, S.; Lertkiatmongkol, P.; Duangkaew, P.; Rongnoparut, P. Modeling of Anopheles minimus Mosquito NADPH-Cytochrome P450 Oxidoreductase (CYPOR) and Mutagenesis Analysis. Int. J. Mol. Sci. 2013, 14, 1788-1801.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics



[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top