The Optical Biosensor Study of Protein-Protein Interactions within Cytochromes P450 Containing Monooxygenase Systems

  • Alexander I. Archakov
  • Yuri D. Ivanov
Part of the Electronics and Biotechnology Advanced (EL.B.A.) Forum Series book series (ELBA, volume 3)


The cytochromes P450 — containing liver microsomal monooxyganase systems play an important role in the oxidation of drugs, toxins, carcinogens, mutagens and other xenobiotics (Archakov and Bachmanova, 1990). It is known that microsomal cytochromes P450 (P450), in particular P450 2B4 (2B4), function by interacting with their redox partners — NADPH-cytochrome P450 reductase (Fp) and cytochrome b5 (b5). Mitochondrial cytochromes P450 (particularly P450scc) hydroxylate cholesterol by acquiring electrons from adrenodoxin (Ad), which in turn accepts them from adrenodoxin reductase (AdR). Similar mechanism operates in the case of bacterial P450s (for example, P450cam), whose electron donor is putidoredoxin (Pd), which acquires electrons from putidoredoxin reductase (PdR). In some studies, protein-protein interactions were explored by the the spin equilibrium shift (Backes et al., 1985; Tamburini et al., 1985; Lapko et al., 1991; Gerber and Sligar, 1994) and the fluorescence quenching techniques (Davydov et al., 1996), based on which the affinities and kinetic constants of P450 complexes were determined. In other studies, kinetic electron transfer rate constants for interprotein electron transfer were reported (Cush et al., 1993; Tamburini et al., 1985; Eyer and Backes, 1992; Kanaeva et al., 1992; Wu et al., 1994; Voznesensky and Shenkman, 1992; Sevrukova et al., 1994; Gerber and Sligar, 1994; Lambeth and Kriengsiri, 1985; Lambeth and Kamin 1977; Nakamura et al., 1994). In present-day molecular interaction researches the ever-widening application is finding the optical biosensor method. It enables to study molecular interactions without protein labeling in real time (Johnsson et al., 1991; Cush et al., 1993; Davies et al., 1994; Yeung et al., 1995).


Complex Formation Incubation Mixture Dissociation Rate Constant Redox Partner Binding Curve 
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Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • Alexander I. Archakov
    • 1
  • Yuri D. Ivanov
    • 1
  1. 1.Institute of Biomedical ChemistryRAMSMoscowRussia

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