TY - JOUR
T1 - Structural engineering of a phage lysin that targets Gram-negative pathogens
AU - Lukacik, Petra
AU - Barnard, Travis J.
AU - Keller, Paul W.
AU - Chaturvedi, Kaveri S.
AU - Seddiki, Nadir
AU - Fairman, James W.
AU - Noinaj, Nicholas
AU - Kirby, Tara L.
AU - Henderson, Jeffrey P.
AU - Steven B, Alasdair C.
AU - Hinnebusch, Joseph
AU - Buchanan, Susan K.
PY - 2012/6/19
Y1 - 2012/6/19
N2 - 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.
AB - 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.
KW - Colicin
KW - Muramidase
KW - Plague
KW - TonB-dependent transport
UR - http://www.scopus.com/inward/record.url?scp=84862514534&partnerID=8YFLogxK
U2 - 10.1073/pnas.1203472109
DO - 10.1073/pnas.1203472109
M3 - Article
C2 - 22679291
AN - SCOPUS:84862514534
SN - 0027-8424
VL - 109
SP - 9857
EP - 9862
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 25
ER -