Biochemical characterisation of Cytochrome P450 oxidoreductase from the cattle tick, Rhipicephalus microplus, highlights potential new acaricide target

Research output: Journal Publications and ReviewsRGC 21 - Publication in refereed journalpeer-review

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Author(s)

  • Adeyinka I Fadahunsi
  • Christopher Kumm
  • Kirsty Graham
  • Adalberto A Pérez de León
  • Felix Guerrero
  • Robert D Finn

Detail(s)

Original languageEnglish
Article number102148
Journal / PublicationTicks and Tick-borne Diseases
Volume14
Issue number3
Online published9 Mar 2023
Publication statusPublished - May 2023

Link(s)

Abstract

Management of the cattle tick, Rhipicephalus microplus, presents a challenge because some populations of this cosmopolitan and economically important ectoparasite are resistant to multiple classes of acaricides. Cytochrome P450 oxidoreductase (CPR) is part of the cytochrome P450 (CYP450) monooxygenases that are involved in metabolic resistance by their ability to detoxify acaricides. Inhibiting CPR, the sole redox partner that transfers electrons to CYP450s, could overcome this type of metabolic resistance. This report represents the biochemical characterisation of a CPR from ticks. Recombinant CPR of R. microplus (RmCPR), minus its N-terminal transmembrane domain, was produced in a bacterial expression system and subjected to biochemical analyses. RmCPR displayed a characteristic dual flavin oxidoreductase spectrum. Incubation with nicotinamide adenine dinucleotide phosphate (NADPH) lead to an increase in absorbance between 500 and 600 nm with a corresponding appearance of a peak absorbance at 340–350 nm indicating functional transfer of electrons between NADPH and the bound flavin cofactors. Using the pseudoredox partner, kinetic parameters for both cytochrome c and NADPH binding were calculated as 26.6 ± 11.4 µM and 7.03 ± 1.8 µM, respectively. The turnover, Kcat, for RmCPR for cytochrome c was calculated as 0.08 s−1 which is significantly lower than the CPR homologues of other species. IC50 (Half maximal Inhibitory Concentration) values obtained for the adenosine analogues 2’, 5’ ADP, 2’- AMP, NADP+and the reductase inhibitor diphenyliodonium were: 140, 82.2, 24.5, and 75.3 µM, respectively. Biochemically, RmCPR resembles CPRs of hematophagous arthropods more so than mammalian CPRs. These findings highlight the potential of RmCPR as a target for the rational design of safer and potent acaricides against R. microplus. © 2023 The Author(s).

Research Area(s)

  • Acaricide target, Enzyme kinetics, P450 oxidoreductase, Rhipicephalus microplus

Citation Format(s)

Biochemical characterisation of Cytochrome P450 oxidoreductase from the cattle tick, Rhipicephalus microplus, highlights potential new acaricide target. / Fadahunsi, Adeyinka I; Kumm, Christopher; Graham, Kirsty et al.
In: Ticks and Tick-borne Diseases, Vol. 14, No. 3, 102148, 05.2023.

Research output: Journal Publications and ReviewsRGC 21 - Publication in refereed journalpeer-review

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