Reference : Properties and crystallization of a genetically engineered, water-soluble derivative of ...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/81280
Properties and crystallization of a genetically engineered, water-soluble derivative of penicillin-binding protein 5 of Escherichia coli K12
English
Ferreira, Luis C [Institut für Entwicklungsbiologie (Tübigen) > Abteilung Biochemie > > >]
Schwarz, Uli [Institut für Entwicklungsbiologie (Tübigen) > Abteilung Biochemie > > >]
Keck, Wolfgang [Institut für Entwicklungsbiologie (Tübigen) > Abteilung Biochemie > > >]
Charlier, Paulette mailto [Université de Liège - ULg > Institut de Physique > Laboratoire de Cristallographie > >]
Dideberg, Otto [Université de Liège - ULg > Institut de Physique > Laboratoire de Cristallographie > >]
Ghuysen, Jean-Marie [Université de Liège - ULg > Institut de Chimie > Service de Microbiologie > >]
15-Jan-1988
European Journal of Biochemistry
Blackwell Science
171
1-2
11-16
Yes (verified by ORBi)
International
0014-2956
1432-1033
Oxford
United Kingdom
[en] bacterial proteins ; carrier proteins/genetics ; crystallization ; escherichia coli/enzymology/*genetics ; genetic engineering ; hexosyltransferases ; molecular weight ; muramoylpentapeptide carboxypeptidase/genetics ; penicillin-binding proteins ; peptidyl transferases ; serine endopeptidases/genetics ; solubility
[en] Derivatives of the Escherichia coli penicillin-binding protein 5 (PBP5) with truncated carboxyl terminals were obtained by altering the carboxyl-coding end of the dacA gene. After cloning the modified dacA gene into a runaway-replication-control plasmid, one clone that overproduced and excreted the desired protein into the periplasm was used as a source for the isolation of a water-soluble PBP5 (i.e. PBP5S). In PBP5S the carboxyl-terminal 21-amino-acid region of the wild-type protein was replaced by a short 9-amino-acid segment. Milligram amounts of PBP5S were purified by penicillin affinity chromatography in the absence of detergents or of chaotropic agents. PBP5S was stable and possessed DD-carboxypeptidase activity without added Triton X-100. Upon reaction with [14C]benzylpenicillin it was converted into a rather short-lived acyl-enzyme complex, as observed with PBP5. Both PBP5 and PBP5S were crystallized. In contrast to PBP5, PBP5S yielded enzymatically active, well-formed prismatic crystals suitable for X-ray analysis.
Fonds de la Recherche Scientifique Médicale - FRSM
Researchers ; Professionals
http://hdl.handle.net/2268/81280
also: http://hdl.handle.net/2268/77983

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