Chemical Probes of Cytochrome P450 Structure

  • R. Bernhardt
Part of the Handbook of Experimental Pharmacology book series (HEP, volume 105)


Chemical modification of proteins is based on the fact that some amino acid side chains of proteins are accessible and, under specific conditions, are reactive enough to bind the probe (for detailed information see, e.g., Means and Feeney 1971). The main problem is of causing alterations of the protein structure when introducing the probe such as: (1) local effects (short-range steric and noncovalent binding interactions, including salt bridges, H-bonds, hydrophobic and van der Waals interactions as well as local conformation changes); (2) global effects (overall changes of conformation or solvation that may affect, or be propagated throughout, the molecule as a whole), and (3) specific long-range effects (these might include specific pathways whereby structural modifications at a site are transmitted to another site) (Ackers and Smith 1985). This is a problem common not only to methods introducing probes, but also to methods changing the primary structure of proteins (de novo and semisynthesis with amino acid exchanges, site-directed mutagenesis). That is why one has to be very careful in performing and interpreting modification studies on proteins. Nevertheless, chemical probes have been widely used to study the structure and function of cytochromes P450 as well as functionally linked protein-protein interactions in cytochrome P450 systems. The broad interest in using this method is due to the fact that the overwhelming number of cytochromes P450 so far isolated are membrane bound (Nebert et al. 1991) and therefore it is difficult to crystallize them so that direct structural information on these cytochromes P450 has so far not been available.


Chemical Probe Succinic Anhydride Cytochrome P450 System Microsomal Cytochrome Adrenodoxin Reductase 
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© Springer-Verlag Berlin Heidelberg 1993

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  • R. Bernhardt

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