The threat of engineered bacterial bioweapons that are resistant to current antibiotic treatments provides a powerful mandate to develop novel classes of therapeutic compounds. This proposal presents a concerted effort of three complementary groups at the Rockefeller University (Dr. Vincent Fischetti, Dr. Alexander Tomasz, and Dr. Erec Stebbins) to identify novel inhibitors of the Category A pathogens, B. anthracis and Y. pestis, as well as Category B food and water safety threats. The first and central research project is based on the observation that resistance to bacteriophage lytic enzymes, such as PlyG for B. anthracis, is an extremely rare event, from which we hypothesize that the binding domain of these enzymes targets a critical component in the bacterial cell wall to assure phage release (in a sense through evolution, the phage have performed the high throughput assay to identify this component). Thus, the pathway to the synthesis of this wall component represents a target for antibiotic development. The core project will therefore focus on the identification, isolation and characterization of the lytic enzyme binding substrate. Once identified, the pathway towards its synthesis will be determined. Those enzymes catalyzing the pathway will be isolated, purified and crystallized, and inhibitors identified. The cell wall structural experience of Alex Tomasz, feeds into the core project but also broadens it to a search for additional novel targets against B. anthracis by high resolution analysis of the cell walls and by the isolation of conditional lethal mutants - using in vivo transposition of a mariner type element. Cell walls occupy center stage in his project since (i) the target of PlyG is in the wall; (ii) cell walls play an important role in the devastating septic shock caused by B. anthracis infections and (iii) the genome of B. anthracis contains wall-related determinants that are virulence factors in other bacteria. Erec Stebbins will provide the expertise in protein structure and crystallization along with the identification of small molecule inhibitors for these structures. His project expands the novel therapeutics search to include Y. pestis and Category B pathogens through a translational component in computation drug discovery coupled to structural determination. In total, these efforts target central virulence factors in critical biowarfare pathogens.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
1U19AI056510-01
Application #
6688188
Study Section
Special Emphasis Panel (ZAI1-GPJ-M (M1))
Program Officer
Baker, Phillip J
Project Start
2003-08-01
Project End
2008-01-31
Budget Start
2003-08-01
Budget End
2004-01-31
Support Year
1
Fiscal Year
2003
Total Cost
$731,685
Indirect Cost
Name
Rockefeller University
Department
Microbiology/Immun/Virology
Type
Other Domestic Higher Education
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
Schuch, Raymond; Pelzek, Adam J; Raz, Assaf et al. (2013) Use of a bacteriophage lysin to identify a novel target for antimicrobial development. PLoS One 8:e60754
Xu, Yong; Brenning, Benjamin; Clifford, Adrianne et al. (2013) Discovery of Novel Putative Inhibitors of UDP-GlcNAc 2-Epimerase as Potent Antibacterial Agents. ACS Med Chem Lett 4:1142-1147
Hu, Xin; Vujanac, Milos; Southall, Noel et al. (2013) Inhibitors of the Yersinia protein tyrosine phosphatase through high throughput and virtual screening approaches. Bioorg Med Chem Lett 23:1056-62
Schmitz, Jonathan E; Daniel, Anu; Collin, Mattias et al. (2008) Rapid DNA library construction for functional genomic and metagenomic screening. Appl Environ Microbiol 74:1649-52
Velloso, Lucas M; Bhaskaran, Shyam S; Schuch, Raymond et al. (2008) A structural basis for the allosteric regulation of non-hydrolysing UDP-GlcNAc 2-epimerases. EMBO Rep 9:199-205
Hu, Xin; Prehna, Gerd; Stebbins, C Erec (2007) Targeting plague virulence factors: a combined machine learning method and multiple conformational virtual screening for the discovery of Yersinia protein kinase A inhibitors. J Med Chem 50:3980-3
Prehna, Gerd; Stebbins, C Erec (2007) A Rac1-GDP trimer complex binds zinc with tetrahedral and octahedral coordination, displacing magnesium. Acta Crystallogr D Biol Crystallogr 63:628-35
Fischetti, Vincent A (2007) In vivo acquisition of prophage in Streptococcus pyogenes. Trends Microbiol 15:297-300
Cheng, Qi; Fischetti, Vincent A (2007) Mutagenesis of a bacteriophage lytic enzyme PlyGBS significantly increases its antibacterial activity against group B streptococci. Appl Microbiol Biotechnol 74:1284-91
Daniel, Anu; Bonnen, Penelope E; Fischetti, Vincent A (2007) First complete genome sequence of two Staphylococcus epidermidis bacteriophages. J Bacteriol 189:2086-100

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