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 two 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 trea Gram- negative infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI081059-09
Application #
9207105
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Ernst, Nancy Lewis
Project Start
2008-12-15
Project End
2019-01-31
Budget Start
2017-02-01
Budget End
2018-01-31
Support Year
9
Fiscal Year
2017
Total Cost
$475,072
Indirect Cost
$187,968
Name
Harvard University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
082359691
City
Cambridge
State
MA
Country
United States
Zip Code
02138
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
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:
Okuda, Suguru; Sherman, David J; Silhavy, Thomas J et al. (2016) Lipopolysaccharide transport and assembly at the outer membrane: the PEZ model. Nat Rev Microbiol 14:337-45
Lee, James; Xue, Mingyu; Wzorek, Joseph S et al. (2016) Characterization of a stalled complex on the ?-barrel assembly machine. Proc Natl Acad Sci U S A 113:8717-22
May, Janine M; Sherman, David J; Simpson, Brent W et al. (2015) Lipopolysaccharide transport to the cell surface: periplasmic transport and assembly into the outer membrane. Philos Trans R Soc Lond B Biol Sci 370:
Simpson, Brent W; May, Janine M; Sherman, David J et al. (2015) Lipopolysaccharide transport to the cell surface: biosynthesis and extraction from the inner membrane. Philos Trans R Soc Lond B Biol Sci 370:
Hagan, Christine L; Wzorek, Joseph S; Kahne, Daniel (2015) Inhibition of the ?-barrel assembly machine by a peptide that binds BamD. Proc Natl Acad Sci U S A 112:2011-6
Maloj?i?, Goran; Andres, Dorothee; Grabowicz, Marcin et al. (2014) LptE binds to and alters the physical state of LPS to catalyze its assembly at the cell surface. Proc Natl Acad Sci U S A 111:9467-72
Sherman, David J; Lazarus, Michael B; Murphy, Lea et al. (2014) Decoupling catalytic activity from biological function of the ATPase that powers lipopolysaccharide transport. Proc Natl Acad Sci U S A 111:4982-7

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