Journal Article

A specific peptide inhibitor of the class B metallo-β-lactamase L-1 from <i>Stenotrophomonas maltophilia</i> identified using phage display

François Sanschagrin and Roger C. Levesque

in Journal of Antimicrobial Chemotherapy

Published on behalf of British Society for Antimicrobial Chemotherapy

Volume 55, issue 2, pages 252-255
Published in print February 2005 | ISSN: 0305-7453
Published online February 2005 | e-ISSN: 1460-2091 | DOI: http://dx.doi.org/10.1093/jac/dkh550
A specific peptide inhibitor of the class B metallo-β-lactamase L-1 from Stenotrophomonas maltophilia identified using phage display

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Objectives: In Gram-negative bacteria, resistance to β-lactam antibiotics and to known inhibitors mediated by metallo-β-lactamases is a major concern and a serious threat to public health. Since no clinically useful inhibitors are available against class B metallo-β-lactamases, the aim of the study was to identify peptides as inhibitors.

Methods: The L-1 metalloenzyme from Stenotrophomonas maltophilia was cloned, over-expressed, purified to homogeneity and used in screening of peptide libraries by phage display with a selective and competitive biopanning assay. This was based upon the high affinity of L-1 for cefoxitin and its slow hydrolysis.

Results: From six peptides, the consensus sequence Cys-Val-His-Ser-Pro-Asn-Arg-Glu-Cys was identified as a promising inhibitor of L-1 hydrolytic activity. This peptide showed a mixed inhibition of L-1 with a Ki competitive of 16 ± 4 μM and a Ki uncompetitive of 9 ± 1 μM. The same peptide was prepared without flanking Cys residues and demonstrated no detectable inhibition of L-1 hydrolytic activity with nitrocefin as a substrate. These data confirmed the importance of the peptide conformation for the inhibition of L-1 hydrolytic activity. Further analysis revealed rescue by Zn2+ ions. The mixed inhibition indicated peptide binding near the active site of L-1 and blocking of zinc atoms for optimal conformation in the pocket of the active site.

Conclusion: This is the first report of a peptide inhibitor for Class B metallo-β-lactamases. It will be used as a lead to identify more potent small molecule inhibitors via peptidomimetics.

Keywords: L-1 β-lactamase; selective biopanning; β-lactamase inhibitors

Journal Article.  2161 words.  Illustrated.

Subjects: Medical Oncology ; Critical Care

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