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Characterization of VanY n , a novel D,D‐peptidase/D,D‐carboxypeptidase involved in glycopeptide antibiotic resistance in Nonomuraea sp. ATCC 39727
Author(s) -
Binda Elisa,
Marcone Giorgia L.,
Pollegioni Loredano,
Marinelli Flavia
Publication year - 2012
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2012.08706.x
Subject(s) - peptidoglycan , glycopeptide , dipeptidase , biochemistry , glycopeptide antibiotic , biology , escherichia coli , carboxypeptidase , peptide sequence , microbiology and biotechnology , gene , antibiotics , enzyme , bacteria , genetics , vancomycin , staphylococcus aureus
VanY n is a novel protein involved in the mechanism of self‐resistance in Nonomuraea sp. ATCC 39727, which produces the glycopeptide antibiotic A40926, the precursor of the second‐generation dalbavancin, which is in phase III of clinical development. VanY n (196 residues) is encoded by the dbv7 gene within the dbv biosynthetic cluster devoted to A40926 production. C‐terminal His6‐tagged VanY n was successfully expressed as a soluble and active protein in Escherichia coli. The analysis of the sequence suggests the presence of a hydrophobic transmembrane portion and two conserved sequences (SxHxxGxAxD and ExxH) in the extracytoplasmic domain that are potentially involved in coordination of Zn 2+ and catalytic activity. The presence of these conserved sequences indicates a similar mechanism of action and substrate binding in VanY n as in VanY, VanX and VanXY Zn 2+ ‐dependent d , d ‐carboxypeptidases and d ‐Ala‐ d ‐Ala dipeptidases acting on peptidoglycan maturation and involved in glycopeptide resistance in pathogens. On substrates mimicking peptidoglycan precursors, VanY n shows d , d ‐carboxypeptidase and d , d ‐dipeptidase activity, but lacks d , d ‐carboxyesterase ability on d ‐Ala‐ d ‐Lac‐terminating peptides. VanY n belongs to the metallo‐ d , d ‐carboxypeptidase family, but it is inhibited by β‐lactams. Its characterization provides new insights into the evolution and transfer of resistance determinants from environmental glycopeptide‐producing actinomycetes (such as Nonomuraea sp.) to glycopeptide‐resistant pathogens (enterococci and staphylococci). It may also contribute to an early warning system for emerging resistance mechanisms following the introduction into clinics of a second‐generation glycopeptide such as dalbavancin. Database
The nucleotide sequence of vanY n is available in the GenBank data base under accession number CAD91202