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 journalArticle

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.

LanguageEnglish
Pages7738-7746
Number of pages9
JournalBiochemistry
Volume44
Issue number21
Early online date7 May 2005
DOIs
Publication statusPublished - 31 May 2005

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Serine-Type D-Ala-D-Ala Carboxypeptidase
Catalytic Domain
Enzyme Inhibitors
X ray crystallography
X Ray Crystallography
Streptomyces
Roofs
Serine
Hydrolysis
Rate constants
Substrates
Enzymes
beta-sultam

Cite this

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
Llinás, Antonio ; Ahmed, Naveed ; Cordaro, Massimiliano ; Laws, Andrew P. ; Frère, Jean Marie ; Delmarcelle, Michael ; Silvaggi, Nicholas R. ; Kelly, Judith A. ; Page, Michael I. / Inactivation of Bacterial DD-Peptidase by β-Sultams. In: Biochemistry. 2005 ; Vol. 44, No. 21. pp. 7738-7746.
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Llinás, A, Ahmed, N, Cordaro, M, Laws, AP, Frère, JM, Delmarcelle, M, Silvaggi, NR, Kelly, JA & Page, MI 2005, 'Inactivation of Bacterial DD-Peptidase by β-Sultams', Biochemistry, vol. 44, no. 21, pp. 7738-7746. https://doi.org/10.1021/bi050110o

Inactivation of Bacterial DD-Peptidase by β-Sultams. / Llinás, Antonio; Ahmed, Naveed; Cordaro, Massimiliano; Laws, Andrew P.; Frère, Jean Marie; Delmarcelle, Michael; Silvaggi, Nicholas R.; Kelly, Judith A.; Page, Michael I.

In: Biochemistry, Vol. 44, No. 21, 31.05.2005, p. 7738-7746.

Research output: Contribution to journalArticle

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AU - Delmarcelle, Michael

AU - Silvaggi, Nicholas R.

AU - Kelly, Judith A.

AU - Page, Michael I.

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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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Llinás A, Ahmed N, Cordaro M, Laws AP, Frère JM, Delmarcelle M et al. Inactivation of Bacterial DD-Peptidase by β-Sultams. Biochemistry. 2005 May 31;44(21):7738-7746. https://doi.org/10.1021/bi050110o