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Chemosensing Ensembles for Monitoring Biomembrane Transport in Real Time
Author(s) -
Ghale Garima,
Lanctôt Adrienne G.,
Kreissl Hannah T.,
Jacob Maik H.,
Weingart Helge,
Winterhalter Mathias,
Nau Werner M.
Publication year - 2014
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201309583
Subject(s) - analyte , chemistry , protamine , biophysics , lipid bilayer , fluorescence , biomolecule , transmembrane protein , surface plasmon resonance , membrane , combinatorial chemistry , biochemistry , nanotechnology , chromatography , biology , nanoparticle , materials science , heparin , physics , receptor , quantum mechanics
The efficacy of drugs and biomolecules relies on their ability to pass through the bilayer. The development of methods to directly and sensitively monitor these membrane transport processes has remained an experimental challenge. A macrocyclic host ( p ‐sulfonatocalix[4]arene or cucurbit[7]uril) and a fluorescent dye (lucigenin or berberine) are encapsulated as a chemosensing ensemble inside liposomes, which allows for a direct, real‐time fluorescence monitoring of the passage of unlabeled bioorganic analytes. This in vitro assay is transferable to different channel proteins and analytes, has potential for fluorescence‐based screening, e.g., of channel modulators, and yields the absolute kinetics of translocation. Using this new biophysical method, we observed for the first time direct rapid translocation of protamine, an antimicrobial peptide, through the bacterial transmembrane protein OmpF.