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Multifunctional basic motif in the glycine receptor intracellular domain induces subunit-specific sorting
Journal article   Open access   Peer reviewed

Multifunctional basic motif in the glycine receptor intracellular domain induces subunit-specific sorting

N Melzer, C Villmann, Kristina Becker, K Harvey, Robert J Harvey, N Vogel, C J Kluck, M Kneussel and C M Becker
Journal of Biological Chemistry, Vol.285(6), pp.3730-3739
2010
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https://doi.org/10.1074/jbc.M109.030460View
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Abstract

nucleus/nuclear import protein/sorting intracellular trafficking membrane proteins receptor structure-function glycine receptor karyopherin multifunctional motif sorting trafficking
The strychnine-sensitive glycine receptor (GlyR) is a ligand-gated ion channel that mediates fast synaptic inhibition in the vertebrate central nervous system. As a member of the family of Cys-loop receptors, it assembles from five homologous subunits (GlyRα1-4 and -β). Each subunit contains an extracellular ligand binding domain, four transmembrane domains (TM), and an intracellular domain, formed by the loop connecting TM3 and TM4 (TM3-4 loop). The TM3-4 loops of the subunits GlyRα1 and -α3 harbor a conserved basic motif, which is part of a potential nuclear localization signal. When tested for functionality by live cell imaging of green fluorescent protein and β-galactosidase-tagged domain constructs, the TM3-4 loops of GlyRα1 and -α3, but not of GlyRα2 and -β, exhibited nuclear sorting activity. Subunit specificity may be attributed to slight amino acid alterations in the basic motif. In yeast two-hybrid screening and GST pulldown assays, karyopherin α3 and α4 were found to interact with the TM3-4 loop, providing a molecular mechanism for the observed intracellular trafficking. These results indicate that the multifunctional basic motif of the TM3-4 loop is capable of mediating a karyopherin-dependent intracellular sorting of full-length GlyRs. © 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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