Abstract

Antibiotic resistant Gram-negative infections pose a serious threat to human health. The outer membrane of Gram-negative bacteria is a unique structure essential for survival;it also functions as a physical barrier to block entry of many classes of antibiotics and thereby render them ineffective. This research is directed towards understanding the structure and function of three multi-protein machines responsible for the biogenesis of two major components of the outer membrane, lipopolysaccharide (LPS) and outer membrane proteins (OMPs). To understand the protein-protein interactions within each machine and their molecular structures, biochemical and structural studies will be undertaken. To dissect the functions of the individual components of these machines, the assembly of LPS and OMPs will be reconstituted in vitro. A better understanding of the protein machinery and the processes in which they are involved may lead to the discovery of inhibitors that could ultimately be developed to treat Gram-negative infections.

Public Health Relevance

The research proposed here is directed towards understanding the protein machinery responsible for the biogenesis of the outer membrane of Gram-negative bacteria, a structure that is essential for their survival. A better understanding of the protein components of this machinery and the processes in which they are involved may lead to the discovery of inhibitors that could ultimately be developed for therapeutic uses to treat Gram- negative infections.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI081059-03
Application #
7993544
Study Section
Synthetic and Biological Chemistry A Study Section (SBCA)
Program Officer
Korpela, Jukka K
Project Start
2008-12-15
Project End
2013-11-30
Budget Start
2010-12-01
Budget End
2011-11-30
Support Year
3
Fiscal Year
2011
Total Cost
$464,721
Indirect Cost

  Institution

Name
Harvard University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
082359691
City
Cambridge
State
MA
Country
United States
Zip Code
02138

  Publications

Bertani, Blake R; Taylor, Rebecca J; Nagy, Emma et al. (2018) A cluster of residues in the lipopolysaccharide exporter that selects substrate variants for transport to the outer membrane. Mol Microbiol 109:541-554
Mandler, Michael D; Baidin, Vadim; Lee, James et al. (2018) Novobiocin Enhances Polymyxin Activity by Stimulating Lipopolysaccharide Transport. J Am Chem Soc 140:6749-6753
Sherman, David J; Xie, Ran; Taylor, Rebecca J et al. (2018) Lipopolysaccharide is transported to the cell surface by a membrane-to-membrane protein bridge. Science 359:798-801
Zhang, Ge; Baidin, Vadim; Pahil, Karanbir S et al. (2018) Cell-based screen for discovering lipopolysaccharide biogenesis inhibitors. Proc Natl Acad Sci U S A 115:6834-6839
Lee, James; Sutterlin, Holly A; Wzorek, Joseph S et al. (2018) Substrate binding to BamD triggers a conformational change in BamA to control membrane insertion. Proc Natl Acad Sci U S A 115:2359-2364
Xie, Ran; Taylor, Rebecca J; Kahne, Daniel (2018) Outer Membrane Translocon Communicates with Inner Membrane ATPase To Stop Lipopolysaccharide Transport. J Am Chem Soc 140:12691-12694
May, Janine M; Owens, Tristan W; Mandler, Michael D et al. (2017) The Antibiotic Novobiocin Binds and Activates the ATPase That Powers Lipopolysaccharide Transport. J Am Chem Soc 139:17221-17224
Moison, Eileen; Xie, Ran; Zhang, Ge et al. (2017) A Fluorescent Probe Distinguishes between Inhibition of Early and Late Steps of Lipopolysaccharide Biogenesis in Whole Cells. ACS Chem Biol 12:928-932
Wzorek, Joseph S; Lee, James; Tomasek, David et al. (2017) Membrane integration of an essential ?-barrel protein prerequires burial of an extracellular loop. Proc Natl Acad Sci U S A 114:2598-2603
Simpson, Brent W; Owens, Tristan W; Orabella, Matthew J et al. (2016) Identification of Residues in the Lipopolysaccharide ABC Transporter That Coordinate ATPase Activity with Extractor Function. MBio 7:

Showing the most recent 10 out of 38 publications