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- W4311876512 endingPage "110" @default.
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- W4311876512 abstract "Some of the most dangerous bacterial pathogens (Gram-negative and mycobacterial) deploy a formidable secondary membrane barrier to reduce the influx of exogenous molecules. For Gram-negative bacteria, this second exterior membrane is known as the outer membrane (OM), while for the Gram-indeterminate Mycobacteria, it is known as the myco membrane. Although different in composition, both the OM and mycomembrane are key structures that restrict the passive permeation of small molecules into bacterial cells. Although it is well-appreciated that such structures are principal determinants of small molecule permeation, it has proven to be challenging to assess this feature in a robust and quantitative way or in complex, infection-relevant settings. Herein, we describe the development of the bacterial chloro-alkane penetration assay (BaCAPA), which employs the use of a genetically encoded protein called HaloTag, to measure the uptake and accumulation of molecules into model Gram-negative and mycobacterial species, Escherichia coli and Mycobacterium smegmatis, respectively, and into the human pathogen Mycobacterium tuberculosis. The HaloTag protein can be directed to either the cytoplasm or the periplasm of bacteria. This offers the possibility of compartmental analysis of permeation across individual cell membranes. Significantly, we also showed that BaCAPA can be used to analyze the permeation of molecules into host cell-internalized E. coli and M. tuberculosis, a critical capability for analyzing intracellular pathogens. Together, our results show that BaCAPA affords facile measurement of permeability across four barriers: the host plasma and phagosomal membranes and the diderm bacterial cell envelope." @default.
- W4311876512 created "2023-01-02" @default.
- W4311876512 creator A5020173783 @default.
- W4311876512 creator A5025721506 @default.
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- W4311876512 creator A5042720391 @default.
- W4311876512 creator A5068275315 @default.
- W4311876512 creator A5078147131 @default.
- W4311876512 creator A5081309101 @default.
- W4311876512 date "2022-12-19" @default.
- W4311876512 modified "2023-09-23" @default.
- W4311876512 title "Measurement of Small Molecule Accumulation into Diderm Bacteria" @default.
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- W4311876512 doi "https://doi.org/10.1021/acsinfecdis.2c00435" @default.
- W4311876512 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/36530146" @default.
- W4311876512 hasPublicationYear "2022" @default.