Human pathogenic gram-negative bacteria, such as Escherichia coli (E. Coli) and Salmonella enterica cause millions of deaths, thousands of fatalities and billions of dollars in medical expenses and lost wages. A critical process that enables these organisms to overcome the hosts immune response is the injection of proteins directly into infected mammalian (host) cells. Some of these injected effectors, function as enzymes that modify the structure/function of the host proteins, which in turn diminish the cells' ability to fight off the infection. One family of these enzymes are N-glycosyltransferases, which appear to play critical, but yet unknown, function in pathogenicity. These enzymes add a single ?-N-acetyl-D-glucosamine (GlcNAc) to the guanidinium groups of arginine residues. The primary focus of this application is to evaluate/develop methodology that will enable the generation of site-specific ?-N-GlcNAc antibodies. We feel that this ability will open the door to the creation of a wide range of Abs that will have a wide-ranging impact the fields of pharmacy, microbiology, and biochemistry.
Public Health Relevance Statement: Human pathogenic gram-negative bacteria, such as Escherichia coli (E. Coli) and Salmonella enterica cause millions of deaths, thousands of fatalities and billions of dollars in medical expenses and lost wages. A critical process that enables these organisms to overcome the hosts immune response is the injection of proteins directly into infected mammalian (host) cells. Some of these injected effectors, function as enzymes that modify the structure/function of the host proteins, which in turn diminish the cells' ability to fight off the infection. One family of these enzymes are N-glycosyltransferases, which appear to play critical, but yet unknown, function in pathogenicity. These enzymes add a single ?-N-acetyl-D-glucosamine (GlcNAc) to the guanidinium groups of arginine residues. The primary focus of this application is to evaluate/develop methodology that will enable the generation of site-specific ?-N-GlcNAc antibodies. We feel that this ability will open the door to the creation of a wide range of Abs that will have a wide-ranging impact the fields of pharmacy, microbiology, and biochemistry.
Project Terms: Affinity; Animal Model; Antibiotic Resistance; Antibiotics; Antibodies; Arginine; Bacteria; Biochemistry; Biological Assay; Cell physiology; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Citrobacter rodentium; combat; Communities; cost; Death Domain; Development; Diarrhea; Economics; Enzymes; Equipment; Escherichia coli; Escherichia coli EHEC; FADD protein; Family; fighting; foodborne; foodborne illness; foodborne outbreak; Future; Generations; Glucosamine; Glyceraldehyde-3-Phosphate Dehydrogenases; glycosyltransferase; Gram-Negative Bacteria; guanidinium; Hemolytic-Uremic Syndrome; Hospitalization; Human; Immune response; Immune system; Immunization; Immunoprecipitation; In Vitro; in vivo; Infection; Inflammatory disease of the intestine; inhibitor/antagonist; Injections; Mammals; Measures; Medical; Methodology; Microbiology; Modification; Monitor; Monoclonal Antibodies; mortality; Mus; Names; novel; Organism; pathogen; Pathogenesis; pathogenic Escherichia coli; Pathogenicity; Pharmacy (field); Phase; Play; Procedures; Process; Protein-Carbohydrate Interaction; Proteins; Public Health; recombinant peptide; Research; Research Personnel; Resistance; Role; Salmonella; Salmonella enterica; Sampling; Site; Specificity; Structure; sugar; Tertiary Protein Structure; TRADD gene; treatment strategy; Tumor Necrosis Factor Receptor; UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase; Virulence; Wages; Western Blotting
