Reference : Importance of the Two Tryptophan Residues in the Streptomyces R61 Exocellular Dd-Peptidase
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/79706
Importance of the Two Tryptophan Residues in the Streptomyces R61 Exocellular Dd-Peptidase
English
Bourguignon-Bellefroid, Catherine [Université de Liège - ULiège > Institut de Chimie > Laboratoire d'Enzymologie / Centre d'Ingénierie des Proteines > > >]
Wilkin, Jean-Marc [Université de Liège - ULiège > Institut de Chimie > Laboratoire d'Enzymologie / Centre d'Ingénierie des Proteines > >]
Joris, Bernard mailto [Université de Liège - ULiège > Institut de Chimie > Laboratoire d'Enzymologie / Centre d'Ingénierie des Proteines > > >]
Aplin, Robin T. [University of Oxford > > The Dyson Perrins Laboratory > >]
Houssier, Claude [University of Oxford > > The Dyson Perrins Laboratory > >]
Prendergast, Franklin G. [Mayo Foundation > Department of Biochemistry and Molecular Biology > > >]
Van Beeumen, Jozef [Rijksuniversiteit-Gent > > > > Laboratorium voor Microbiologie en Microbiele Genetica > >]
Ghuysen, Jean-Marie [Université de Liège - ULiège > Institut de Chimie > Laboratoire d'Enzymologie / Centre d'Ingenierie des Proteines/Laboratoire de Chimie Macromoleculaire et Chimie Physique, > > >]
Frère, Jean-Marie mailto [Université de Liège - ULiège > Institut de Chimie > Centre d'ingénierie des protéines > > >]
1-Mar-1992
Biochemical Journal
Portland Press
282
Pt 2
361-367
Yes (verified by ORBi)
International
0264-6021
1470-8728
London
United Kingdom
[en] Streptomyces ; Peptidase ; Tryptophan ; Site directed mutagenesis ; β-Lactamase ; Enzymatic activity ; Molecular microenvironment ; Chemical modification ; Kinetic parameter ; Streptomycetaceae ; Actinomycetales ; Actinomycetes ; Bacteria ; Enzyme
[fr] Tryptophane ; Mutagenèse dirigée ; Activité enzymatique ; Microenvironnement moléculaire ; Modification chimique ; Paramètre cinétique ; Bactérie
[es] Triptófano ; Mutagénesis dirigida ; Actividad enzimática ; Microambiente molecular ; Modificación química ; Parámetro cinético
[en] Modification of the Streptomyces R61 DD-peptidase by N-bromosuccinimide resulted in a rapid loss of enzyme activity. In consequence, the role of the enzyme's two tryptophan residues was investigated by site-directed mutagenesis. Trp271 was replaced by Leu. The modification yielded a stable enzyme whose structural and catalytic properties were similar to those of the wild-type protein. Thus the Trp271 residue, though almost invariant among the beta-lactamases of classes A and C and the low-Mr penicillin-binding proteins, did not appear to be essential for enzyme activity. Mutations of the Trp233 into Leu and Ser strongly decreased the enzymic activity, the affinity for beta-lactams and the protein stability. Surprisingly, the benzylpenicilloyl-(W233L)enzyme deacylated at least 300-fold more quickly than the corresponding acyl-enzyme formed with the wild-type protein and gave rise to benzylpenicilloate instead of phenylacetylglycine. This mutant DD-peptidase thus behaved as a weak beta-lactamase.
Centre d'Ingénierie des Protéines - CIP
Fonds de la Recherche Scientifique Médicale - FRSM ; Fonds de la Recherche Scientifique (Communauté française de Belgique) - F.R.S.-FNRS ; Intercommunale du Réseau Social d'Insertion et d'Accueil - IRSIA
Researchers ; Professionals
http://hdl.handle.net/2268/79706

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