Continuous spectrofluorometric and spectrophotometric assays for NADPH-cytochrome P450 reductase activity using 5-cyano-2,3-ditolyl tetrazolium chloride.

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  • Author(s): Kim DH;Kim DH; Yim SK; Kim KH; Ahn T; Yun CH
  • Source:
    Biotechnology letters [Biotechnol Lett] 2009 Feb; Vol. 31 (2), pp. 271-5. Date of Electronic Publication: 2008 Oct 21.
  • Publication Type:
    Journal Article; Research Support, Non-U.S. Gov't
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Kluwer Academic Publishers Country of Publication: Netherlands NLM ID: 8008051 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1573-6776 (Electronic) Linking ISSN: 01415492 NLM ISO Abbreviation: Biotechnol Lett Subsets: MEDLINE
    • Publication Information:
      Publication: 1999- : Dordrecht : Kluwer Academic Publishers
      Original Publication: [Kew, Eng., Science and Technology Letters]
    • Subject Terms:
      NADPH-Ferrihemoprotein Reductase/*chemistry ; Spectrometry, Fluorescence/*methods ; Tetrazolium Salts/*analysis ; Tetrazolium Salts/*chemistry; Enzyme Activation ; Enzyme Stability ; NADPH-Ferrihemoprotein Reductase/analysis ; Reproducibility of Results ; Sensitivity and Specificity ; Substrate Specificity
    • Abstract:
      Mammalian NADPH-cytochrome P450 reductase (CPR) transfers electrons from NADPH to cytochrome P450 enzymes and other several microsomal enzymes. It also catalyzes the one-electron reduction of many chemicals and drugs. Reduction of 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) by CPR was assessed as a method for monitoring CPR activity. The electrons released from NADPH by CPR were transferred to CTC in the reaction medium, and CTC reduction activity could be assessed spectrophotometrically and spectrofluorometrically. The reduction kinetics of CTC follows classical Michaelis-Menten kinetics (K(m) = 50 microM, k(cat) = 2,520 min(-1)). This method offers a continuous assay of the enzymatic activity of CPR.
    • Accession Number:
      0 (Tetrazolium Salts)
      102568-47-8 (5-cyano-2,3-ditolyltetrazolium)
      EC 1.6.2.4 (NADPH-Ferrihemoprotein Reductase)
    • Publication Date:
      Date Created: 20081023 Date Completed: 20090202 Latest Revision: 20081231
    • Publication Date:
      20231215
    • Accession Number:
      10.1007/s10529-008-9865-4
    • Accession Number:
      18941714