Allosteric effects on substrate dissociation from cytochrome P450 3A4 in nanodiscs observed by ensemble and single-molecule fluorescence spectroscopy

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

Actions (login required)

Administrator's edit/view item Administrator's edit/view item