title: Compounds targeting disulfide bond forming enzyme DsbB of Gram-negative bacteria.
abstract: In bacteria, disulfide bonds confer stability on many proteins exported to the cell envelope or beyond. These proteins include numerous bacterial virulence factors, and thus bacterial enzymes that promote disulfide bond formation represent targets for compounds inhibiting bacterial virulence. Here, we describe a new target- and cell-based screening methodology for identifying compounds that inhibit the disulfide bond-forming enzymes Escherichia coli DsbB (EcDsbB) or Mycobacterium tuberculosis VKOR (MtbVKOR), which can replace EcDsbB, although the two are not homologs. Initial screening of 51,487 compounds yielded six specifically inhibiting EcDsbB. These compounds share a structural motif and do not inhibit MtbVKOR. A medicinal chemistry approach led us to select related compounds, some of which are much more effective DsbB inhibitors than those found in the screen. These compounds inhibit purified DsbB and prevent anaerobic growth of E. coli. Furthermore, these compounds inhibit all but one of the DsbBs of nine other Gram-negative pathogenic bacteria tested.
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Cristina
authors: C
authors: Landeta
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Jessica L
authors: JL
authors: Blazyk
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Feras
authors: F
authors: Hatahet
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Brian M
authors: BM
authors: Meehan
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Markus
authors: M
authors: Eser
authors: Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, USA.
authors: Alissa
authors: A
authors: Myrick
authors: Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
authors: Ludmila
authors: L
authors: Bronstain
authors: Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, USA.
authors: Shoko
authors: S
authors: Minami
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Holly
authors: H
authors: Arnold
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Na
authors: N
authors: Ke
authors: Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, USA.
authors: Eric J
authors: EJ
authors: Rubin
authors: Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
authors: Barbara C
authors: BC
authors: Furie
authors: Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
authors: Bruce
authors: B
authors: Furie
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Jon
authors: J
authors: Beckwith
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Rachel
authors: R
authors: Dutton
authors: Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Dana
authors: D
authors: Boyd
mesh_terms: D000362:Agar / Q000737:chemistry; D000900:Anti-Bacterial Agents / Q000737:chemistry; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D020134:Catalytic Domain; D002626:Chemistry, Pharmaceutical / Q000379:methods; D020650:Combinatorial Chemistry Techniques; D004220:Disulfides; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004579:Electron Transport; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D008826:Microbial Sensitivity Tests; D020102:Mycobacterium smegmatis / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D011487:Protein Conformation; D019704:Protein Disulfide-Isomerases / Q000037:antagonists & inhibitors / Q000737:chemistry; D011550:Pseudomonas aeruginosa / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004220:Disulfides; C079503:DsbB protein, Bacteria; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D000362:Agar; D019704:Protein Disulfide-Isomerases; C526648:dsbA protein, E coli