<Emphasis Type="Italic">LAPTM4A</Emphasis> interacts with <Emphasis Type="Italic">hOCT2</Emphasis> and regulates its endocytotic recruitment |
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| Authors: | A Grabner S Brast S Sucic S Bierer B Hirsch H Pavenstädt H H Sitte E Schlatter G Ciarimboli |
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| Institution: | 1.Medizinische Klinik und Poliklinik D, Abteilung für Experimentelle Nephrologie,Universit?tsklinikum Münster,Münster,Germany;2.Center for Physiology and Pharmacology, Institute of Pharmacology,Medical University Vienna,Vienna,Austria;3.Klinik und Poliklinik für Urologie,Universit?tsklinikum Münster,Münster,Germany |
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| Abstract: | Human organic cation transporter 2 (hOCT2) is involved in the transport of endogenous and exogenous organic cations mainly in cells of the kidney and the brain. Here,
we focus on the regulation of hOCT2 by direct protein–protein interaction. Screening within a mating-based split-ubiquitin-yeast-two-hybrid system (mBSUS) revealed
the lysosomal-associated protein transmembrane 4 alpha (LAPTM4A) as a potential interacting protein. Interaction of LAPTM4A and hOCT2 was confirmed by pulldown assays, FRET microscopy analysis and immunofluorescence microscopy. Functionally, overexpression
of LAPTM4A significantly decreased ASP+ uptake in HEK293 cells stably transfected with hOCT2, suggesting a negative regulation of hOCT2-mediated transport. Furthermore, overexpression of LAPTM4A leads to a significantly decreased hOCT2 plasma membrane expression in surface biotinylation experiments. In addition, significant expression of LAPTM4A in human kidney was demonstrated by immunoblotting and immunofluorescence. |
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