Cytochrome P450 are an enzyme superfamily with a diverse range of functions and are particularly interesting in pharmaceutical, bioanalytical and environmental applications. They are able to metabolise drugs, degrade toxins and catalyse reactions for valuable products. Looking for new substrates which are active with P450 is a very active area of research however determining if a new substrate has reacted isn’t always easy.
There are known substrates for Cytochrome P450 BM3 which do not induce a spin shift on the heme iron, making the reaction undetectable by UV-vis. Expensive and time consuming assays with cofactors and LC-MS studies is one option but Gianfranco Gilardi et al. have developed a fluorescence probe which indicates binding for both physiological and non-physiological substrates and can be used to determine dissociation constants.
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Fluorescence detection of ligand binding to labeled cytochrome P450 BM3
Valentina E. V. Ferrero, Giovanna Di Nardo, Gianluca Catucci, Sheila J. Sadeghi and Gianfranco Gilardi
Dalton Trans., 2012, Advance Article