Article (Scientific journals)
Primary structure of the Streptomyces R61 extracellular DD-peptidase. 2. Amino acid sequence data
Joris, Bernard; Jacques, Philippe; Frère, Jean-Marie et al.
1987In European Journal of Biochemistry, 162 (3), p. 519-524
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Keywords :
amino acid sequence; amino acids/analysis; binding sites; cyanogen bromide; hydrolysis; kinetics; muramoylpentapeptide carboxypeptidase/antagonists & inhibitors/*isolation; purification; penicillanic acid; peptide fragments/isolation & purification; phthalic acids; streptomyces/*enzymology; sulfhydryl compounds/analysis; trypsin
Abstract :
[en] In order to confirm the Streptomyces codon usage, the Streptomyces R61 DD-peptidase was fragmented by cyanogen bromide cleavage of the carboxymethylated protein, trypsin digestion of the carboxymethylated protein and trypsin digestion of the protein treated with beta-iodopenicillinate and endoxo-delta 4-tetrahydrophthalic acid. The isolated peptides, which altogether represented more than 50% of the polypeptide chain, were sequenced. The data thus obtained were in excellent agreement with the primary structure of the protein as deduced from the nucleotide sequence of the cloned gene. Though a weak acylating agent, beta-iodopenicillanate reacted selectively with the active site of the DD-peptidase and formed an adduct which mas much more stable than that formed with benzylpenicillin, thus facilitating the isolation and characterization of the active-site peptide.
Disciplines :
Microbiology
Biochemistry, biophysics & molecular biology
Author, co-author :
Joris, Bernard ;  Université de Liège - ULiège > Faculté de Médecine > Service de Microbiologie appliquée aux sciences pharmaceutiques
Jacques, Philippe ;  Université de Liège - ULiège > Faculté de Médecine > Service de Microbiologie appliquée aux sciences pharmaceutiques
Frère, Jean-Marie ;  Université de Liège - ULiège > Facultée de Médecine > Service de Microbiologie appliquée aux sciences pharmaceutiques
Ghuysen, Jean-Marie ;  Université de Liège - ULiège > Faculté de Médecine > Service de Microbiologie appliquée aux sciences pharmaceutiques
Van Beeumen, J.;  Rijksuniversiteit-Gent > Laboratorium voor Microbiologie
Language :
English
Title :
Primary structure of the Streptomyces R61 extracellular DD-peptidase. 2. Amino acid sequence data
Publication date :
1987
Journal title :
European Journal of Biochemistry
ISSN :
0014-2956
eISSN :
1432-1033
Publisher :
Blackwell Science, Oxford, United Kingdom
Volume :
162
Issue :
3
Pages :
519-524
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE]
FRSM - Fonds de la Recherche Scientifique Médicale [BE]
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since 03 February 2011

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