Nath, A., Koo, P. K., Rhoades, E., Atkins, W. M. (November 2008) Allosteric effects on substrate dissociation from cytochrome P450 3A4 in nanodiscs observed by ensemble and single-molecule fluorescence spectroscopy. J Am Chem Soc, 130 (47). pp. 15746-7. ISSN 0002-7863
Abstract
Cytochrome P450 (CYP) 3A4 is a major human drug-metabolizing enzyme and displays pharmacologically relevant allosteric kinetics caused by multiple substrate and/or effector binding. Here, in the first single-molecule (SM) fluorescence studies of CYPs, we use total internal reflection fluorescence microscopy to measure residence times of the fluorescent dye Nile Red in CYP3A4 incorporated in surface-immobilized lipid Nanodiscs, with and without the effector alpha-naphthoflavone. We find direct evidence that CYP3A4 effectors can decrease substrate off-rates, providing a possible mechanism for effector-mediated enhancement of substrate metabolism. These interesting results highlight the potential of SM methods in studies of CYP allosteric mechanisms.
| Item Type: | Paper |
|---|---|
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes |
| CSHL Authors: | |
| Communities: | CSHL labs > Koo Lab |
| Depositing User: | Matthew Dunn |
| Date: | 26 November 2008 |
| Date Deposited: | 16 Sep 2019 18:53 |
| Last Modified: | 16 Sep 2019 18:53 |
| PMCID: | PMC2649694 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/38386 |
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