Assays for Analyzing the Role of Transport Proteins in the Uptake and the Vectorial Transport of Substances Affecting Cell Viability

Journal article


Publication Details

Author(s): Taghikhani EP, Fromm M, König J
Journal: Methods in molecular biology (Clifton, N.J.)
Publication year: 2017
Volume: 1601
Pages range: 123-135
ISSN: 1064-3745


Abstract

Endogenous compounds, drugs, or other xenobiotics may affect cell viability. A prerequisite for intracellular cell damage is the uptake of such substances across the plasma membrane into cells. Furthermore, the subsequent transporter-mediated export out of cells may influence cell viability. Therefore, transport proteins mediating the uptake (uptake transporter) or export (export pumps) of substances in and out of cells are important determinants of cell viability. Uptake transporters mostly belong to the superfamily of solute carriers (SLC transporters), whereas export pumps are members of the ABC-transporter superfamily (ATP-binding cassette). Cell systems recombinantly overexpressing uptake transporters (single transfectants) or multiple-transfected cell models expressing simultaneously an uptake transporter together with an export pump (double transfectants) are important in vitro tools for analyzing protein-mediated transport of potentially cell toxic compounds.Here we describe different in vitro transport assays for the functional analysis of transport proteins. Using single-transfected HEK293 cells stably overexpressing an uptake transporter, substances can be tested as potential substrates (uptake assay) or potential transport inhibitors (inhibition assay) for the respective transport protein. Vectorial transport of substances with the uptake across the basolateral plasma membrane and the export across the apical membrane of polarized grown MDCKII cells can be analyzed using double-transfected cell models with the simultaneous overexpression of an uptake transport and an export pump in vectorial transport assays, thereby mimicking physiological transport processes, e.g., in liver or kidney.


FAU Authors / FAU Editors

Fromm, Martin Prof. Dr.
Lehrstuhl für Klinische Pharmakologie und Klinische Toxikologie
König, Jörg Prof. Dr.
Lehrstuhl für Klinische Pharmakologie und Klinische Toxikologie
Taghikhani, Emir Puyan
Lehrstuhl für Klinische Pharmakologie und Klinische Toxikologie


How to cite

APA:
Taghikhani, E.P., Fromm, M., & König, J. (2017). Assays for Analyzing the Role of Transport Proteins in the Uptake and the Vectorial Transport of Substances Affecting Cell Viability. Methods in molecular biology (Clifton, N.J.), 1601, 123-135. https://dx.doi.org/10.1007/978-1-4939-6960-9_11

MLA:
Taghikhani, Emir Puyan, Martin Fromm, and Jörg König. "Assays for Analyzing the Role of Transport Proteins in the Uptake and the Vectorial Transport of Substances Affecting Cell Viability." Methods in molecular biology (Clifton, N.J.) 1601 (2017): 123-135.

BibTeX: 

Last updated on 2018-07-10 at 02:27