Reference : Catalytic mechanism of the Streptomyces K15 DD-transpeptidase/penicillin-binding prot...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/3971
Catalytic mechanism of the Streptomyces K15 DD-transpeptidase/penicillin-binding protein probed by site-directed mutagenesis and structural analysis
English
Rhazi, Noureddine mailto [Université de Liège - ULg > > Centre d'ingénierie des protéines >]
Charlier, Paulette mailto [Université de Liège - ULg > > Centre d'ingénierie des protéines >]
Dehareng, Dominique mailto [Université de Liège - ULg > > Centre d'ingénierie des protéines >]
Engher, Danièle [> > > >]
Vermeire, Marcel [> > > >]
Frère, Jean-Marie mailto [Université de Liège - ULg > Département des sciences de la vie > Département des sciences de la vie >]
Nguyen-Disteche, Martine [> > > >]
Fonze, Eveline [> > > >]
18-Mar-2003
Biochemistry
Amer Chemical Soc
42
10
2895-2906
Yes (verified by ORBi)
International
0006-2960
Washington
[en] penicillin binding protein ; K15 transpeptidase ; site-directed mutagenesis
[fr] structural analysis
[en] The Streptomyces K15 penicillin-binding DD-transpeptidase is presumed to be involved in peptide cross-linking during bacterial cell wall peptidoglycan assembly. To gain insight into the catalytic mechanism, the roles of residues Lys38, Ser96, and Cys98, belonging to the structural elements defining the active site cleft, have been investigated by site-directed mutagenesis, biochemical studies, and X-ray diffraction analysis. The Lys38His and Ser96Ala mutations almost completely abolished the penicillin binding and severely impaired the transpeptidase activities while the geometry of the active site was essentially the same as in the wild-type enzyme. It is proposed that Lys38 acts as the catalytic base that abstracts a proton from the active serine Ser35 during nucleophilic attack and that Ser96 is a key intermediate in the proton transfer from the Ogamma of Ser35 to the substrate leaving group nitrogen. The role of these two residues should be conserved among penicillin-binding proteins containing the Ser-Xaa-Asn/Cys sequence in motif 2. Conversion of Cys98 into Asn decreased the transpeptidase activity and increased hydrolysis of the thiolester substrate and the acylation rate with most beta-lactam antibiotics. Cys98 is proposed to play the same role as Asn in motif 2 of other penicilloyl serine transferases in properly positioning the substrate for the catalytic process.
Politique Scientifique Fédérale (Belgique) = Belgian Federal Science Policy
Researchers ; Professionals ; Students
http://hdl.handle.net/2268/3971

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