[en] Penicillin-binding protein 4a (PBP4a) from Bacillus subtilis was overproduced and purified to homogeneity. It clearly exhibits DD-carboxypeptidase and thiolesterase activities in vitro. Although highly isologous to the Actinomadura sp. strain R39 DD-peptidase (B. Granier, C. Duez, S. Lepage, S. Englebert, J. Dusart, O. Dideberg, J. van Beeumen, J. M. Frere, and J. M. Ghuysen, Biochem. J. 282:781-788, 1992), which is rapidly inactivated by many beta-lactams, PBP4a is only moderately sensitive to these compounds. The second-order rate constant (k(2)/K) for the acylation of the essential serine by benzylpenicillin is 300,000 M(-1) s(-1) for the Actinomadura sp. strain R39 peptidase, 1,400 M(-1) s(-1) for B. subtilis PBP4a, and 7,000 M(-1) s(-1) for Escherichia coli PBP4, the third member of this class of PBPs. Cephaloridine, however, efficiently inactivates PBP4a (k(2)/K = 46,000 M(-1) s(-1)). PBP4a is also much more thermostable than the R39 enzyme.
Research Center/Unit :
CIP - Centre d'Ingénierie des Protéines - ULiège
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Duez, Colette ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Vanhove, Marc; Université de Liège - ULiège > Centre d'Ingénierie des Protéines > 2001
Gallet, Xavier; Faculté Universitaire des Sciences Agronomiques de Gembloux - FUSAGx > Laboratopire de Biophysique Moléculaire Numérique > 2001
Bouillenne, Fabrice ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Docquier, Jean-Denis ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Brans, Alain ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Frère, Jean-Marie ; Université de Liège - ULiège > Département des sciences de la vie > Enzymologie
Language :
English
Title :
Purification and Characterization of Pbp4a, a New Low-Molecular-Weight Penicillin-Binding Protein from Bacillus subtilis
Alternative titles :
[fr] Purification et caractérisation du PBP4a, un nouveau PBP de faible poids moléculaire, de Bacillus subtilis
Publication date :
March 2001
Journal title :
Journal of Bacteriology
ISSN :
0021-9193
eISSN :
1098-5530
Publisher :
American Society for Microbiology, Washington, United States - District of Columbia
Volume :
183
Issue :
5
Pages :
1595-9
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
Purification et caractérisation du PBP4a de Bacillus subtilis
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique BELSPO - Politique scientifique fédérale
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