Lookup NU author(s): Ally Vitali,
Dr James Stach
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Antisense PNA oligomers targeting essential genes (acpP or ftsZ) and conjugated to the delivery peptide L((KFF)3K) show complete growth inhibition of wild type E. coli strain (MG1655) with submicromolar MIC. In this study we show that resistant mutants generated against such PNA-peptide conjugates had disruptions in the region of sbmA, a gene encoding an inner membrane peptide transporter. The wild type sensitivity to the PNA conjugates was re-established in the resistance mutants by complementation with sbmA. Furthermore, deletion of sbmA in E. coli AS19, a strain that is sensitive to unmodified PNA, resulted in resistance to PNA. Finally, PNA conjugated with the corresponding non-biological H-D((KFF)3K) peptide retained antibacterial activity in sbmA deletion strains, whereas the same conjugate with a protease-sensitive linker did not. These results clearly identify SbmA as a carrier of naked PNA over the inner bacterial membrane and thereby infer that the peptide is transporting the PNA conjugates over the outer membrane. Strains lacking SbmA were used to screen novel peptide-PNA carriers that were SbmA-independent. Four such PNA-peptide conjugates, H-D((KFF)3K), H-(RFR)4-Ahx-βAla, H-(R-Ahx-R)4-Ahx-βAla, and H-(R-Ahx)6-βAla, were identified that utilize an alternative uptake mechanism but retain their antimicrobial potency. In addition SbmA is the first protein identified to recognize PNA.
Author(s): Ghosal A, Vitali A, Stach JEM, Nielsen PE
Publication type: Article
Publication status: Published
Journal: ACS Chemical Biology
Print publication date: 08/11/2012
ISSN (print): 1554-8929
ISSN (electronic): 1554-8937
Publisher: American Chemical Society
PubMed id: 23138594
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