Cleavage Specificity of Enterococcus faecalis EnpA (EF1473), a Peptidoglycan Endopeptidase Related to the LytM/Lysostaphin Family of Metallopeptidases - Normandie Université Accéder directement au contenu
Article Dans Une Revue (Data Paper) Journal of Molecular Biology Année : 2010

Cleavage Specificity of Enterococcus faecalis EnpA (EF1473), a Peptidoglycan Endopeptidase Related to the LytM/Lysostaphin Family of Metallopeptidases

Résumé

Enterococcus faecalis EnpA (EF1473) is a 1721-residue predicted protein encoded by prophage 03 that displays similarity to the staphylolytic glycyl– glycyl endopeptidases lysostaphin and LytM. We purified a catalytically active fragment of the protein, EnpAC, comprising residues 1374–1505 and showed that the recombinant polypeptide efficiently cleaved cross-linked muropeptides generated by muramidases, but was poorly active in intact sacculi. Analysis of the products of digestion of purified dimers by mass spectrometry indicated that EnpAC cleaves the D-Ala-L-Ala bond formed by the D,D-transpeptidase activity of penicillin-binding proteins in the last cross-linking step of peptidoglycan synthesis. Synthetic D was identified as the minimum substrate of EnpAC indicating that interaction of the enzyme with the donor peptide stem of cross-linked dimers is sufficient for its activity. Peptidoglycan was purified from various bacterial species and digested with mutanolysin and EnpAC to assess enzyme specificity. EnpAC did not cleave direct cross-links, but tolerated extensive variation in crossbridges with respect to both their length (one to five residues) and their amino acid sequence. Recognition of the donor stem of cross-linked dimers could account for the substrate specificity of EnpAC, which is significantly broader in comparison to endopeptidases belonging to the lysostaphin family.

Dates et versions

hal-04157426 , version 1 (10-07-2023)

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Citer

François Reste de Roca, Caroline Duché, Shengli Dong, Alain Rincé, Lionel Dubost, et al.. Cleavage Specificity of Enterococcus faecalis EnpA (EF1473), a Peptidoglycan Endopeptidase Related to the LytM/Lysostaphin Family of Metallopeptidases. Journal of Molecular Biology, 2010, 398 (4), pp.507-517. ⟨10.1016/j.jmb.2010.03.033⟩. ⟨hal-04157426⟩
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