In vitro Assay to Evaluate Cation Transport of Ionophores
Huriye D Uzun1,2, Melissa Vázquez-Hernández3, Julia E Bandow3
1Department of Molecular Biochemistry, Faculty of Chemistry and Biochemistry, Ruhr University Bochum, Bochum, Germany.
Abstract:
Ion homeostasis is a fundamental regulator of cellular processes and depends upon lipid membranes, which function as ion permeability barriers. Ionophores facilitate ion transport across cell membranes and offer a way to manipulate cellular ion composition. Here, we describe a calcein quenching assay based on large unilamellar vesicles that we used to evaluate divalent cation transport of the ionophore 4-Br-A23187. This assay can be used to study metal transport by ionophores and membrane proteins, under well-defined conditions. This protocol was validated in: Proteomics (2022), DOI: 10.1002/pmic.202200061 Graphical abstract.
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