Inactivation of Bacterial DD-Peptidase by β-Sultams

Antonio Llinás; Naveed Ahmed; Massimiliano Cordaro; Andrew P. Laws; Jean Marie Frère; Michael Delmarcelle; Nicholas R. Silvaggi; Judith A. Kelly; Michael I. Page

doi:10.1021/bi050110o

Inactivation of Bacterial DD-Peptidase by β-Sultams

Antonio Llinás, Naveed Ahmed, Massimiliano Cordaro, Andrew P. Laws, Jean Marie Frère, Michael Delmarcelle, Nicholas R. Silvaggi, Judith A. Kelly, Michael I. Page

Research output: Contribution to journal › Article

29 Citations (Scopus)

Abstract

N-Acyl-β-sultams are time-dependent, irreversible active site-directed inhibitors of Streptomyces R61 DD-peptidase. The rate of inactivation is first order with respect to β-sultam concentration, and the second-order rate constants show a dependence on pH similar to that for the hydrolysis of a substrate. Inactivation is due to the formation of a stable 1:1 enzyme-inhibitor complex as a result of the active site serine being sulfonylated by the β-sultam as shown by ESI-MS analysis and by X-ray crystallography. A striking feature of the sulfonyl enzyme is that the inhibitor is not bound to the oxyanion hole but interacts extensively with the "roof" of the active site where the Arg 285 is located.

Original language	English
Pages (from-to)	7738-7746
Number of pages	9
Journal	Biochemistry
Volume	44
Issue number	21
Early online date	7 May 2005
DOIs	https://doi.org/10.1021/bi050110o
Publication status	Published - 31 May 2005

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Llinás, A., Ahmed, N., Cordaro, M., Laws, A. P., Frère, J. M., Delmarcelle, M., ... Page, M. I. (2005). Inactivation of Bacterial DD-Peptidase by β-Sultams. Biochemistry, 44(21), 7738-7746. https://doi.org/10.1021/bi050110o

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abstract = "N-Acyl-β-sultams are time-dependent, irreversible active site-directed inhibitors of Streptomyces R61 DD-peptidase. The rate of inactivation is first order with respect to β-sultam concentration, and the second-order rate constants show a dependence on pH similar to that for the hydrolysis of a substrate. Inactivation is due to the formation of a stable 1:1 enzyme-inhibitor complex as a result of the active site serine being sulfonylated by the β-sultam as shown by ESI-MS analysis and by X-ray crystallography. A striking feature of the sulfonyl enzyme is that the inhibitor is not bound to the oxyanion hole but interacts extensively with the {"}roof{"} of the active site where the Arg 285 is located.",

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T1 - Inactivation of Bacterial DD-Peptidase by β-Sultams

AU - Llinás, Antonio

AU - Ahmed, Naveed

AU - Cordaro, Massimiliano

AU - Laws, Andrew P.

AU - Frère, Jean Marie

AU - Delmarcelle, Michael

AU - Silvaggi, Nicholas R.

AU - Kelly, Judith A.

AU - Page, Michael I.

PY - 2005/5/31

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N2 - N-Acyl-β-sultams are time-dependent, irreversible active site-directed inhibitors of Streptomyces R61 DD-peptidase. The rate of inactivation is first order with respect to β-sultam concentration, and the second-order rate constants show a dependence on pH similar to that for the hydrolysis of a substrate. Inactivation is due to the formation of a stable 1:1 enzyme-inhibitor complex as a result of the active site serine being sulfonylated by the β-sultam as shown by ESI-MS analysis and by X-ray crystallography. A striking feature of the sulfonyl enzyme is that the inhibitor is not bound to the oxyanion hole but interacts extensively with the "roof" of the active site where the Arg 285 is located.

AB - N-Acyl-β-sultams are time-dependent, irreversible active site-directed inhibitors of Streptomyces R61 DD-peptidase. The rate of inactivation is first order with respect to β-sultam concentration, and the second-order rate constants show a dependence on pH similar to that for the hydrolysis of a substrate. Inactivation is due to the formation of a stable 1:1 enzyme-inhibitor complex as a result of the active site serine being sulfonylated by the β-sultam as shown by ESI-MS analysis and by X-ray crystallography. A striking feature of the sulfonyl enzyme is that the inhibitor is not bound to the oxyanion hole but interacts extensively with the "roof" of the active site where the Arg 285 is located.

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10.1021/bi050110o

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