Abstract
P450s have attracted tremendous attention owing to not only their involvement in the metabolism of drug molecules and endogenous substrates but also the unusual nature of the reaction they catalyze, namely, the oxidation of unactivated C–H bonds. The binding of substrates to P450s, which is usually viewed as the first step in the catalytic cycle, has been studied extensively via a variety of biochemical and biophysical approaches. These studies were directed towards answering different questions related to P450s, including mechanism of oxidation, substrate properties, unusual substrate oxidation kinetics, function, and active-site features. Some of the substrate binding studies extending over a period of more than 40 years of dedicated work have been summarized in this review and categorized by the techniques employed in the binding studies.
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Notes
The term “absorbance silent” refers to a substrate–P450 interaction that does not involve a change in heme Soret spectra and therefore cannot be detected by absorption spectroscopy.
The term “thermodynamic signatures” is used by the authors to describe plots characterizing the thermodynamic parameters (free energy, enthalpy, and entropy) associated with the binding of a particular ligand.
Abbreviations
- αNF:
-
α-naphthoflavone
- CD:
-
circular dichroism
- EPR:
-
electron paramagnetic resonance
- FRET:
-
Förster resonance energy transfer
- HSQC:
-
heteronuclear single quantum coherence
- ITC:
-
isothermal titration calorimetry
- SAM:
-
self-assembled monolayer
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F.P.G. is suported by US Public Health Service grants R37 CA090426 and P30 ES000267. We thank W. Comstock for editorial assistance.
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Isin, E.M., Guengerich, F.P. Substrate binding to cytochromes P450. Anal Bioanal Chem 392, 1019–1030 (2008). https://doi.org/10.1007/s00216-008-2244-0
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DOI: https://doi.org/10.1007/s00216-008-2244-0