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2017 | 77 | Suppl.1 |
Tytuł artykułu

Trafficking of SLC6A14 – An amino acid transporter B(0,+) to plasma membrane

Warianty tytułu
Języki publikacji
EN
Abstrakty
EN
INTRODUCTION: The solute carrier 6 (SLC6) family of genes codes transporters for neurotransmitters, amino acids, osmolytes and energy metabolites. In the brain SLC6 transporters play an important role in the reuptake of GABA, serotonin, dopamine, noradrenaline, glycine and proline. SLC6A14 is a Na/Cl-dependent amino acid transporter ATB(0,+) – specific towards neutral and cationic amino acids. It is present in astrocytes and in the blood-brain barrier. In the first step of trafficking to plasma membrane – ER exit, the neurotransmitter transporters interact with specific isoforms of SEC24 proteins, the components of Coatomer II (COPII). AIM(S): To verify, which proteins are involved in trafficking of ATB(0,+) to the cell surface from ER. METHOD(S): Trafficking of rat SLC6A14 to plasma membrane was studied in heterologous expression system. RESULTS: Immunofluorescence analysis showed that SLC6A14 appears in the plasma membrane after 48 h, with the majority of the transporter not leaving ER. This observation was confirmed by biotinylation of surface proteins and analysis of SLC6A14 glycosylation status in order to distinguish core glycosylation (taking place in ER) from full glycosylation (taking place in Golgi apparatus). Trafficking was attenuated after co‑transfection with the dominant negative mutants of Sar1 GTPase – the first protein involved in COPII formation. Further studies demonstrated that out of four SEC24 proteins, exclusively SEC24C co-localized and interacted with SLC6A14, a result confirmed in the proximity ligation assay. CONCLUSIONS: These observations confirm a hypothesis that lysine in position +2 towards an ER export signal is crucial forspecific interaction of SLC6 transporters with SEC24C. FINANCIAL SUPPORT: This work has been supported by the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement no 665735 (Bio4Med) and by the funding from Polish Ministry of Science and Higher Education within 2016–2020 funds for the implementation of international projects (agreement no 3548/H2020/COFUND2016/2).
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-
Rocznik
Tom
77
Numer
Opis fizyczny
p.111
Twórcy
autor
  • Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology of Polish Academy of Sciences, Warsaw, Poland
  • Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology of Polish Academy of Sciences, Warsaw, Poland
autor
  • Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology of Polish Academy of Sciences, Warsaw, Poland
autor
  • Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology of Polish Academy of Sciences, Warsaw, Poland
autor
  • Center for Physiology and Pharmacology, Institute of Pharmacology, Medical University of Vienna, Vienna, Austria
  • Center for Physiology and Pharmacology, Institute of Pharmacology, Medical University of Vienna, Vienna, Austria
autor
  • Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology of Polish Academy of Sciences, Warsaw, Poland
Bibliografia
Typ dokumentu
Bibliografia
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