Structural engineering of a phage lysin that targets Gram-negative pathogens

Petra Lukacik, Travis J. Barnard, Paul W. Keller, Kaveri S. Chaturvedi, Nadir Seddiki, James W. Fairman, Nicholas Noinaj, Tara L. Kirby, Jeffrey P. Henderson, Alasdair C. Steven B, Joseph Hinnebusch, Susan K. Buchanan

Research output: Contribution to journalArticlepeer-review

87 Scopus citations

Abstract

Bacterial pathogens are becoming increasingly resistant to antibiotics. As an alternative therapeutic strategy, phage therapy reagents containing purified viral lysins have been developed against Gram-positive organisms but not against Gram-negative organisms due to the inability of these types of drugs to cross the bacterial outer membrane. We solved the crystal structures of a Yersinia pestis outer membrane transporter called FyuA and a bacterial toxin called pesticin that targets this transporter. FyuA is a β-barrel membrane protein belonging to the family of TonB dependent transporters, whereas pesticin is a soluble protein with two domains, one that binds to FyuA and another that is structurally similar to phage T4 lysozyme. The structure of pesticin allowed us to design a phage therapy reagent comprised of the FyuA binding domain of pesticin fused to the N-terminus of T4 lysozyme. This hybrid toxin kills specific Yersinia and pathogenic E. coli strains and, importantly, can evade the pesticin immunity protein (Pim) giving it a distinct advantage over pesticin. Furthermore, because FyuA is required for virulence and is more common in pathogenic bacteria, the hybrid toxin also has the advantage of targeting primarily disease-causing bacteria rather than indiscriminately eliminating natural gut flora.

Original languageEnglish
Pages (from-to)9857-9862
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume109
Issue number25
DOIs
StatePublished - Jun 19 2012

Keywords

  • Colicin
  • Muramidase
  • Plague
  • TonB-dependent transport

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