Visualization and Quantification of Transmembrane Ion Transport into Giant Unilamellar Vesicles

Transmembrane ion transporters (ionophores) are widely investigated as supramolecular agents with potential for biological activity. Tests are usually performed in synthetic membranes that are assembled into large unilamellar vesicles (LUVs). However transport must be followed through bulk propertie...

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Bibliographic Details
Published in:Angewandte Chemie 2015-02, Vol.127 (7), p.2165-2169
Main Authors: Valkenier, Hennie, López Mora, Néstor, Kros, Alexander, Davis, Anthony P.
Format: Article
Language:eng ; ger
Subjects:
Anionen
Bulk transporters
Chemistry
Cystic fibrosis
Encapsulation
Fluorescence
Ion transport
Ionentransport
Membranen
Microscopy
Riesige unilamellare Vesikel
Supramolekulare Chemie
Transport
Vesicles
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Summary:Transmembrane ion transporters (ionophores) are widely investigated as supramolecular agents with potential for biological activity. Tests are usually performed in synthetic membranes that are assembled into large unilamellar vesicles (LUVs). However transport must be followed through bulk properties of the vesicle suspension, because LUVs are too small for individual study. An alternative approach is described whereby ion transport can be revealed and quantified through direct observation. The method employs giant unilamellar vesicles (GUVs), which are 20–60 μm in diameter and readily imaged by light microscopy. This allows characterization of individual GUVs containing transporter molecules, followed by studies of transport through fluorescence emission from encapsulated indicators. The method provides new levels of certainty and relevance, given that the GUVs are similar in size to living cells. It has been demonstrated using a highly active anion carrier, and should aid the development of compounds for treating channelopathies such as cystic fibrosis. Wahre Größe: Der Ionentransport durch Membranen wird üblicherweise an Vesikeln studiert, die zu klein für eine Beobachtung mit Routinemikroskopie sind. Der Einsatz riesiger unilamellarer Vesikel ermöglicht nun eine solche direkte Visualisierung. Bei einer Untersuchung des Chlorid‐Nitrat‐Austauschs mithilfe eines starken Anionophors konnte der Anionentransport in zellgroße Membrankompartimente quantifiziert werden.
ISSN:0044-8249
1521-3757
DOI:10.1002/ange.201410200