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Localization of Cytochrome P450 in Membranes: Reconstituted Systems

  • D. Müller-Enoch
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Part of the Handbook of Experimental Pharmacology book series (HEP, volume 105)

Abstract

Since the successful isolation of the components of the cytochrome P450 dependent monooxygenase system by Lu and Coon (1968), their reconstitution to an enzymatically active system was established by the use of phosphatidylcholines by Strobel et al. (1970). Among various acyl derivatives of glycerol-3-phosphorylcholine the dioleoyl-derivative was most effective. The most commonly used and convenient system for reconstitution in numerous studies over the years and until now for routine work consists of the appropriate purified cytochrome P450 (0.3–1µM), a 1.5 molar excess of purified NADPH-cytochrome P450 reductase (reductase) and 30 µM l-α-dilauroylglyceryl-3-phosphatidylcholine (DLPC). This system was described by Guengerich et al. (1982) for eight different rat liver cytochrome P450 forms for use with numerous substrates. Generally the enzymatic reaction is started by the addition of NADPH or an NADPH-regenerating system. This simple reconstitution system has been found to be very reproducible, so that data from different laboratories can often be easily compared if the same incubation conditions have been used.

Keywords

Monooxygenase Activity Cytochrome P4502B1 Reductase Complex Zwitterionic Detergent Cytochrome P4S0 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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© Springer-Verlag Berlin Heidelberg 1993

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  • D. Müller-Enoch

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