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

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 {beta}-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.

Authors:
; ; ; ; ; ; ; ; ; ; ;  [1];  [2]
  1. (NIH)
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
OTHERNIHNIAID
OSTI Identifier:
1045041
Resource Type:
Journal Article
Journal Name:
Proc. Natl. Acad. Sci. USA
Additional Journal Information:
Journal Volume: 109; Journal Issue: (25) ; 06, 2012; Journal ID: ISSN 1091-6490
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; ANTIBIOTICS; BACTERIA; CRYSTAL STRUCTURE; DESIGN; IMMUNITY; LYSOZYME; MEMBRANE PROTEINS; MEMBRANES; PATHOGENS; PROTEINS; STRAINS; TARGETS; THERAPY; TOXINS; TRANSPORT; VIRULENCE

Citation Formats

Lukacik, Petra, Barnard, Travis J., Keller, Paul W., Chaturvedi, Kaveri S., Seddiki, Nadir, Fairman, James W., Noinaj, Nicholas, Kirby, Tara L., Henderson, Jeffrey P., Steven, Alasdair C., Hinnebusch, B. Joseph, Buchanan, Susan K., and WU-MED). Structural engineering of a phage lysin that targets Gram-negative pathogens. United States: N. p., 2012. Web. doi:10.1073/pnas.1203472109.
Lukacik, Petra, Barnard, Travis J., Keller, Paul W., Chaturvedi, Kaveri S., Seddiki, Nadir, Fairman, James W., Noinaj, Nicholas, Kirby, Tara L., Henderson, Jeffrey P., Steven, Alasdair C., Hinnebusch, B. Joseph, Buchanan, Susan K., & WU-MED). Structural engineering of a phage lysin that targets Gram-negative pathogens. United States. doi:10.1073/pnas.1203472109.
Lukacik, Petra, Barnard, Travis J., Keller, Paul W., Chaturvedi, Kaveri S., Seddiki, Nadir, Fairman, James W., Noinaj, Nicholas, Kirby, Tara L., Henderson, Jeffrey P., Steven, Alasdair C., Hinnebusch, B. Joseph, Buchanan, Susan K., and WU-MED). Tue . "Structural engineering of a phage lysin that targets Gram-negative pathogens". United States. doi:10.1073/pnas.1203472109.
@article{osti_1045041,
title = {Structural engineering of a phage lysin that targets Gram-negative pathogens},
author = {Lukacik, Petra and Barnard, Travis J. and Keller, Paul W. and Chaturvedi, Kaveri S. and Seddiki, Nadir and Fairman, James W. and Noinaj, Nicholas and Kirby, Tara L. and Henderson, Jeffrey P. and Steven, Alasdair C. and Hinnebusch, B. Joseph and Buchanan, Susan K. and WU-MED)},
abstractNote = {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 {beta}-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.},
doi = {10.1073/pnas.1203472109},
journal = {Proc. Natl. Acad. Sci. USA},
issn = {1091-6490},
number = (25) ; 06, 2012,
volume = 109,
place = {United States},
year = {2012},
month = {11}
}