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Proc Natl Acad Sci U S A. 2018 Jun 26;115(26):6834-6839. doi: 10.1073/pnas.1804670115. Epub 2018 May 7.

Cell-based screen for discovering lipopolysaccharide biogenesis inhibitors.

Author information

1
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138.
2
Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138.
3
California Institute for Biomedical Research, La Jolla, CA 92037.
4
California Institute for Biomedical Research, La Jolla, CA 92037; schultz@scripps.edu txm50@psu.edu kahne@chemistry.harvard.edu.
5
Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115; schultz@scripps.edu txm50@psu.edu kahne@chemistry.harvard.edu.
6
Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, PA 16802.
7
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138; schultz@scripps.edu txm50@psu.edu kahne@chemistry.harvard.edu.
8
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.

Abstract

New drugs are needed to treat gram-negative bacterial infections. These bacteria are protected by an outer membrane which prevents many antibiotics from reaching their cellular targets. The outer leaflet of the outer membrane contains LPS, which is responsible for creating this permeability barrier. Interfering with LPS biogenesis affects bacterial viability. We developed a cell-based screen that identifies inhibitors of LPS biosynthesis and transport by exploiting the nonessentiality of this pathway in Acinetobacter We used this screen to find an inhibitor of MsbA, an ATP-dependent flippase that translocates LPS across the inner membrane. Treatment with the inhibitor caused mislocalization of LPS to the cell interior. The discovery of an MsbA inhibitor, which is universally conserved in all gram-negative bacteria, validates MsbA as an antibacterial target. Because our cell-based screen reports on the function of the entire LPS biogenesis pathway, it could be used to identify compounds that inhibit other targets in the pathway, which can provide insights into vulnerabilities of the gram-negative cell envelope.

KEYWORDS:

ABC transporter; Acinetobacter; LPS biogenesis; MsbA inhibitor; high-throughput screening

Comment in

PMID:
29735709
PMCID:
PMC6042065
DOI:
10.1073/pnas.1804670115
[Indexed for MEDLINE]
Free PMC Article

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