title: A metabolic intermediate of the fructose-asparagine utilization pathway inhibits growth of a Salmonella fraB mutant.
abstract: Insertions in the Salmonella enterica fra locus, which encodes the fructose-asparagine (F-Asn) utilization pathway, are highly attenuated in mouse models of inflammation (>1000-fold competitive index). Here, we report that F-Asn is bacteriostatic to a fraB mutant (IC50 19 μM), but not to the wild-type or a fra island deletion mutant. We hypothesized that the presence of FraD kinase and absence of FraB deglycase causes build-up of a toxic metabolite: 6-phosphofructose-aspartate (6-P-F-Asp). We used biochemical assays to assess FraB and FraD activities, and mass spectrometry to confirm that the fraB mutant accumulates 6-P-F-Asp. These results, together with our finding that mutants lacking fraD or the fra island are not attenuated in mice, suggest that the extreme attenuation of a fraB mutant stems from 6-P-F-Asp toxicity. Salmonella FraB is therefore an excellent drug target, a prospect strengthened by the absence of the fra locus in most of the gut microbiota.
authors.affiliation: Department of Microbial Infection and Immunity, The Ohio State University, Columbus, OH 43210, USA.|Center for Microbial Interface Biology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Anice
authors.initials: A
authors.lastname: Sabag-Daigle
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Henry M
authors.initials: HM
authors.lastname: Blunk
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Anindita
authors.initials: A
authors.lastname: Sengupta
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Jikang
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Bogard
authors.affiliation: Center for Microbial Interface Biology, The Ohio State University, Columbus, OH 43210, USA.|Department of Microbiology, The Ohio State University, Columbus, OH 43210, USA.|Department of Medical Microbiology and Immunology, Faculty of Medicine, Mansoura University, Mansoura, Egypt.
authors.forename: Mohamed M
authors.initials: MM
authors.lastname: Ali
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Stahl
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Vicki H
authors.initials: VH
authors.lastname: Wysocki
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Venkat
authors.initials: V
authors.lastname: Gopalan
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Behrman
authors.affiliation: Department of Microbial Infection and Immunity, The Ohio State University, Columbus, OH 43210, USA.|Center for Microbial Interface Biology, The Ohio State University, Columbus, OH 43210, USA.|Department of Microbiology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Brian M M
authors.initials: BM
authors.lastname: Ahmer
mesh_terms: D000818:Animals; D001216:Asparagine / Q000378:metabolism*; D001224:Aspartic Acid / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D005632:Fructose / Q000378:metabolism*; D005636:Fructosephosphates / Q000494:pharmacology*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D013058:Mass Spectrometry; D055432:Metabolomics; D051379:Mice; D009154:Mutation; D010770:Phosphotransferases; D019779:Salmonella enterica / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*
chemical_list: D001426:Bacterial Proteins; D005636:Fructosephosphates; D005632:Fructose; D001224:Aspartic Acid; D001216:Asparagine; D010770:Phosphotransferases