Surface-enhanced Raman scattering as a tool to probe cytochrome P450-catalysed substrate oxidation
Surface-enhanced Raman scattering was used as a spectroscopic tool to investigate the changes brought upon cytochrome P450BSß after fatty acid binding. Differences in the spectra of substrate-free and substrate-bound enzyme were observed indicating the potential for this method to be used in the screening of P450 substrates. In particular, the binding characteristics of myristic acid, an inherent substrate, and hydroxylauric acid, a product of fatty acid oxidation, towards P450BSß in the presence of H2O2 were investigated. Specific spectral changes could be assigned to changes in the heme environment only for myristic acid, indicating an occurrence of oxidation process characteristic for the enzymatic substrate.
KeywordsSurface-enhanced Raman spectroscopy Bioanalytical methods Protein binding
We gratefully acknowledge financial support through the Federal Ministry of Education and Research (BMBF) grant “Markerfreie Zelldiagnostik mit Nanometerauflösung” (0312032A and 0312032B). This work was supported in part by the Zentrum für Angewandte Chemische Genomik (ZACG), a joint research initiative founded by the European Union and the Ministry of Innovation and Research of the state North Rhine-Westphalia. CMN thanks Max Planck Society for financial support of a Max Planck Fellow research group at the Max Planck Institute of Molecular Physiology, Dortmund. LF was supported by Marie Curie International Incoming Fellowship (project 514582).
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