Article (Scientific journals)
Unexpected tricovalent binding mode of boronic acids within the active site of a penicillin binding protein.
Zervosen, Astrid; Herman, Raphaël; Kerff, Frédéric et al.
2011In Journal of the American Chemical Society
Peer Reviewed verified by ORBi
 

Files


Full Text
Sauvage_JACS_2011.pdf
Publisher postprint (4.49 MB)
Download

All documents in ORBi are protected by a user license.

Send to



Details



Abstract :
[en] Boronic acids bearing appropriate side chains are good inhibitors of serine amidohydrolases. The boron usually adopts a tetrahedral conformation, bound to the nucleophilic serine of the active site and mimicking the transition state of the enzymatic reaction. We have solved the structures of complexes of a penicillin-binding protein, the DD-peptidase from Actinomadura sp. R39, with four amidomethylboronic acids (2,6 dimethoxybenzamidomethylboronic acid, phenylacetamidomethylboronic acid, 2-chlorobenzamidomethylboronic acid, and 2-nitrobenzamidomethylboronic acid) and the pinacol ester derived from phenylacetamidomethylboronic acid. We found that, in each case, the boron forms a tricovalent adduct with Ogamma of Ser49, Ser298, and the terminal amine group of Lys410, three key residues involved in the catalytic mechanism of penicillin-binding proteins. This represents the first tricovalent enzyme-inhibitor adducts observed by crystallography. In two of the five R39-boronate structures, the boronic acid is found as a tricovalent adduct in two monomers of the asymmetric unit and as a monocovalent adduct with the active serine in the two remaining monomers of the asymmetric unit. Formation of the tricovalent complex from a classical monocovalent complex may involve rotation around the Ser49 Calpha-Cbeta bond to place the boron in a position to interact with Ser298 and Lys410, and a twisting of the side chain amide such that its carbonyl oxygen is able to hydrogen bond to the oxyanion hole NH of Thr413. Biphasic kinetics were observed in three of the five cases and details of the reaction between R39 and 2,6-dimethoxybenzamidomethylboronic acid were studied. Observation of biphasic kinetics was not, however, thought to be correlated to formation of tricovalent complexes, assuming that the latter do form in solution. Based on the crystallographic and kinetic results, a reaction scheme for this unexpected inhibition by boronic acids is proposed.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Zervosen, Astrid ;  Université de Liège - ULiège > Centre de recherches du cyclotron
Herman, Raphaël ;  Université de Liège - ULiège > Centre d'ingénierie des protéines
Kerff, Frédéric  ;  Université de Liège - ULiège > Centre d'ingénierie des protéines
Herman, A.
Bouillez, André ;  Université de Liège - ULiège > Département de chimie (sciences) > Département de chimie (sciences)
Prati, F.
Pratt, R. F.
Frère, Jean-Marie ;  Université de Liège - ULiège > Centre d'ingénierie des protéines
Joris, Bernard ;  Université de Liège - ULiège > Département des sciences de la vie > Physiologie et génétique bactériennes
Luxen, André ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie organique de synthèse
Charlier, Paulette ;  Université de Liège - ULiège > Centre d'ingénierie des protéines
Sauvage, Eric 
Language :
English
Title :
Unexpected tricovalent binding mode of boronic acids within the active site of a penicillin binding protein.
Publication date :
16 May 2011
Journal title :
Journal of the American Chemical Society
ISSN :
0002-7863
eISSN :
1520-5126
Publisher :
American Chemical Society, Washington, United States - District of Columbia
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 28 July 2011

Statistics


Number of views
153 (19 by ULiège)
Number of downloads
682 (10 by ULiège)

Scopus citations®
 
35
Scopus citations®
without self-citations
30
OpenCitations
 
35

Bibliography


Similar publications



Contact ORBi