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 #
2R01AI081059-06A1
Application #
8696032
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Korpela, Jukka K
Project Start
2008-12-15
Project End
2019-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
6
Fiscal Year
2014
Total Cost
$457,829
Indirect Cost
$169,171
Name
Harvard University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
082359691
City
Cambridge
State
MA
Country
United States
Zip Code
02138
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Sherman, David J; Okuda, Suguru; Denny, William A et al. (2013) Validation of inhibitors of an ABC transporter required to transport lipopolysaccharide to the cell surface in Escherichia coli. Bioorg Med Chem 21:4846-51
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Ricci, Dante P; Hagan, Christine L; Kahne, Daniel et al. (2012) Activation of the Escherichia coli ?-barrel assembly machine (Bam) is required for essential components to interact properly with substrate. Proc Natl Acad Sci U S A 109:3487-91
Freinkman, Elizaveta; Chng, Shu-Sin; Kahne, Daniel (2011) The complex that inserts lipopolysaccharide into the bacterial outer membrane forms a two-protein plug-and-barrel. Proc Natl Acad Sci U S A 108:2486-91
Hagan, Christine L; Kahne, Daniel (2011) The reconstituted Escherichia coli Bam complex catalyzes multiple rounds of ýý-barrel assembly. Biochemistry 50:7444-6
Chimalakonda, Gitanjali; Ruiz, Natividad; Chng, Shu-Sin et al. (2011) Lipoprotein LptE is required for the assembly of LptD by the beta-barrel assembly machine in the outer membrane of Escherichia coli. Proc Natl Acad Sci U S A 108:2492-7

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