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Multiple substrate binding by cytochrome P450 3A4

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Multiple substrate binding by cytochrome P450 3A4 : Estimation of the number of bound substrate molecules. / Kapelyukh, Yury; Paine, Mark; Marechal, Jean-Didier; Sutcliffe, Michael J.; Wolf, C. Roland; Roberts, Gordon C. K.

In: Drug Metabolism and Disposition, Vol. 36, No. 10, 10.2008, p. 2136-2144.

Research output: Contribution to journalArticle

Harvard

Kapelyukh, Y, Paine, M, Marechal, J-D, Sutcliffe, MJ, Wolf, CR & Roberts, GCK 2008, 'Multiple substrate binding by cytochrome P450 3A4: Estimation of the number of bound substrate molecules' Drug Metabolism and Disposition, vol 36, no. 10, pp. 2136-2144., 10.1124/dmd.108.021733

APA

Kapelyukh, Y., Paine, M., Marechal, J-D., Sutcliffe, M. J., Wolf, C. R., & Roberts, G. C. K. (2008). Multiple substrate binding by cytochrome P450 3A4: Estimation of the number of bound substrate molecules. Drug Metabolism and Disposition, 36(10), 2136-2144. 10.1124/dmd.108.021733

Vancouver

Kapelyukh Y, Paine M, Marechal J-D, Sutcliffe MJ, Wolf CR, Roberts GCK. Multiple substrate binding by cytochrome P450 3A4: Estimation of the number of bound substrate molecules. Drug Metabolism and Disposition. 2008 Oct;36(10):2136-2144. Available from: 10.1124/dmd.108.021733

Author

Kapelyukh, Yury; Paine, Mark; Marechal, Jean-Didier; Sutcliffe, Michael J.; Wolf, C. Roland; Roberts, Gordon C. K. / Multiple substrate binding by cytochrome P450 3A4 : Estimation of the number of bound substrate molecules.

In: Drug Metabolism and Disposition, Vol. 36, No. 10, 10.2008, p. 2136-2144.

Research output: Contribution to journalArticle

Bibtex - Download

@article{b0529b3e2fce4f9eb6ce928f5d5b1d08,
title = "Multiple substrate binding by cytochrome P450 3A4: Estimation of the number of bound substrate molecules",
keywords = "DRUG-DRUG INTERACTIONS, CYP3A4 ACTIVE-SITE, IN-VITRO, LIGAND-BINDING, ASPARTATE-TRANSCARBAMYLASE, KINETICS, COOPERATIVITY, INHIBITION, TESTOSTERONE, MIDAZOLAM",
author = "Yury Kapelyukh and Mark Paine and Jean-Didier Marechal and Sutcliffe, {Michael J.} and Wolf, {C. Roland} and Roberts, {Gordon C. K.}",
year = "2008",
doi = "10.1124/dmd.108.021733",
volume = "36",
number = "10",
pages = "2136--2144",
journal = "Drug Metabolism and Disposition",
issn = "0090-9556",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Multiple substrate binding by cytochrome P450 3A4

T2 - Estimation of the number of bound substrate molecules

A1 - Kapelyukh,Yury

A1 - Paine,Mark

A1 - Marechal,Jean-Didier

A1 - Sutcliffe,Michael J.

A1 - Wolf,C. Roland

A1 - Roberts,Gordon C. K.

AU - Kapelyukh,Yury

AU - Paine,Mark

AU - Marechal,Jean-Didier

AU - Sutcliffe,Michael J.

AU - Wolf,C. Roland

AU - Roberts,Gordon C. K.

PY - 2008/10

Y1 - 2008/10

N2 - <p>Cytochrome P450 3A4, a major drug-metabolizing enzyme in man, is well known to show non-Michaelis-Menten steady-state kinetics for a number of substrates, indicating that more than one substrate can bind to the enzyme simultaneously, but it has proved difficult to obtain reliable estimates of exactly how many substrate molecules can bind. We have used a simple method involving studies of the effect of large inhibitors on the Hill coefficient to provide improved estimates of substrate stoichiometry from simple steady-state kinetics. Using a panel of eight inhibitors, we show that at least four molecules of the widely used CYP3A4 substrate 7-benzyloxyquinoline can bind simultaneously to the enzyme. Computational docking studies show that this is consistent with the recently reported crystal structures of the enzyme. In the case of midazolam, which shows simple Michaelis-Menten kinetics, the inhibitor effects demonstrate that two molecules must bind simultaneously, consistent with earlier evidence, whereas for diltiazem, the experiments provide no evidence for the binding of more than one molecule. The consequences of this "inhibitor-induced cooperativity" for the prediction of pharmacokinetics and drug-drug interactions are discussed.</p>

AB - <p>Cytochrome P450 3A4, a major drug-metabolizing enzyme in man, is well known to show non-Michaelis-Menten steady-state kinetics for a number of substrates, indicating that more than one substrate can bind to the enzyme simultaneously, but it has proved difficult to obtain reliable estimates of exactly how many substrate molecules can bind. We have used a simple method involving studies of the effect of large inhibitors on the Hill coefficient to provide improved estimates of substrate stoichiometry from simple steady-state kinetics. Using a panel of eight inhibitors, we show that at least four molecules of the widely used CYP3A4 substrate 7-benzyloxyquinoline can bind simultaneously to the enzyme. Computational docking studies show that this is consistent with the recently reported crystal structures of the enzyme. In the case of midazolam, which shows simple Michaelis-Menten kinetics, the inhibitor effects demonstrate that two molecules must bind simultaneously, consistent with earlier evidence, whereas for diltiazem, the experiments provide no evidence for the binding of more than one molecule. The consequences of this "inhibitor-induced cooperativity" for the prediction of pharmacokinetics and drug-drug interactions are discussed.</p>

KW - DRUG-DRUG INTERACTIONS

KW - CYP3A4 ACTIVE-SITE

KW - IN-VITRO

KW - LIGAND-BINDING

KW - ASPARTATE-TRANSCARBAMYLASE

KW - KINETICS

KW - COOPERATIVITY

KW - INHIBITION

KW - TESTOSTERONE

KW - MIDAZOLAM

U2 - 10.1124/dmd.108.021733

DO - 10.1124/dmd.108.021733

M1 - Article

JO - Drug Metabolism and Disposition

JF - Drug Metabolism and Disposition

SN - 0090-9556

IS - 10

VL - 36

SP - 2136

EP - 2144

ER -

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