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 | |
| Publication status | Published - 31 May 2005 |
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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 journal › Article
TY - JOUR
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
Y1 - 2005/5/31
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.
UR - http://www.scopus.com/inward/record.url?scp=19644380904&partnerID=8YFLogxK
U2 - 10.1021/bi050110o
DO - 10.1021/bi050110o
M3 - Article
C2 - 15909988
AN - SCOPUS:19644380904
VL - 44
SP - 7738
EP - 7746
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 21
ER -