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High-throughput functional assay for BAM-mediated protein folding in a native membrane

Research Project
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01.02.2020
 - 31.01.2021

For the development of novel antibiotics against Gram-negative bacteria, targets in the outer membrane or of prime interest. There exist two generally essential such targets, the outer membrane protein insertase BAM with the core unit BamA and the LPS insertion machinery LptD with its associated proteins. Two novel classes of inhibitors for BamA, a synthetic as well as a natural antibiotic, were recently discovered (Luther et al, Nature 2019, Imai et al, Nature 2019). This validity of BAM as a target creates an imperative requirement of an assay to screen for further inhibitors of BAM catalytic activity in vitro . While assays with synthetic vesicles have been reported, so far, no assay for studying BAM-catalyzed OMP folding in the native membrane is available. A quantitative assay to study OMP folding in the natural bacterial membrane has recently been developed by teh Hiller lab in Biozentrum, University of Basel. The assay makes use of outer membrane vesicles (OMVs) that are released from Gram-negative bacteria as spherical buds from the outer membrane, and hence contain the lipid composition and distribution of the outer membrane. They thus provide native and physiological conditions to study the OMP biogenesis mechanism. It is the goal of this collaborative project to establish the assay for high-throughput screening. The assay is expected to allow screening of novel antibiotics, as well as hit validation and lead expansion in high-throughput format.

Funding

High-throughput functional assay for BAM-mediated protein folding in a native membrane

Industriegelder, Sonstige Forschungskooperationen (GrantsTool), 02.2020-01.2021 (12)
PI : Hiller, Sebastian.

Members (1)

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Sebastian Hiller

Principal Investigator