Assays for Analyzing the Role of Transport Proteins in the Uptake and the Vectorial Transport of Substances Affecting Cell Viability.
Assays for Analyzing the Role of Transport Proteins in the Uptake and the Vectorial Transport of Substances Affecting Cell Viability.
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分析转运蛋白在影响细胞活力的物质的摄取和载体转运中的作用的测定
DOI：
10.1007/978-1-4939-6960-9_11
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发表时间：
2017
影响因子：
--
通讯作者：
König J.
中科院分区：
文献类型：
--
作者：
Taghikhani E;Fromm M.F.;König J.
内源性化合物、药物或其他外源物质可能会影响细胞的活力。细胞内损伤的一个先决条件是这种物质通过质膜进入细胞。此外，随后的转运蛋白介导的细胞外输出可能会影响细胞的存活。因此，介导细胞内外物质摄取(摄取转运体)或输出(输出泵)的转运蛋白是细胞活力的重要决定因素。摄取转运体大多属于溶质载体超家族(SLC转运体)，而出口泵是ABC转运体超家族(ATP结合盒)的成员。重组过表达摄取转运体的细胞系统(单个转染体)或同时表达一个摄取转运体和一个输出泵的多个转染体细胞模型(双转染体)是分析蛋白质介导的潜在细胞毒性化合物转运的重要体外工具。利用稳定过表达摄取转运蛋白的单一转基因HEK293细胞，可以测试物质作为各自转运蛋白的潜在底物(摄取试验)或潜在运输抑制物(抑制试验)。极化生长的MDCKII细胞具有跨基侧质膜的摄取和跨顶膜的输出的物质的载体运输可以使用在载体运输分析中同时过表达摄取运输和输出泵的双转基因细胞模型来分析，从而模拟例如在肝脏或肾脏的生理运输过程。
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.
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影响因子：
6
作者：
Ciarimboli, Giuliano;Deuster, Dirk;Schlatter, Eberhard
通讯作者：
Schlatter, Eberhard
影响因子：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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