Molecular cloning and partial functional characterization of Tsha3 - a novel modulatory potassium channel alpha-subunit of trout CNS

Autor(en): Piwowarski, T
Panofen, F
Jeserich, G
Stichwörter: BETA-SUBUNITS; bony fishes; CELL RT-PCR; electrophysiology; EXPRESSION; IDENTIFICATION; INACTIVATION; K+ CHANNELS; KV2.1; molecular structure; Neurosciences; Neurosciences & Neurology; regulatory subunits; SHAKER; Shaker-type channels; SODIUM; VOLTAGE-GATED POTASSIUM
Erscheinungsdatum: 2004
Volumen: 124
Ausgabe: 2
Startseite: 124
Seitenende: 133
A novel Shaker-related potassium channel subunit termed Tsha3 that is widely expressed in the CNS of trout was PCR-cloned and sequenced: its deduced amino acid sequence showed an extended N-terminal domain with a high proportion of negatively charged residues and possessed highest similarity with KCNA10, a human epithelial potassium channel. Upon heterologous expression in Sf21 cells, homomeric Tsha3 did not yield voltage-activated potassium channels but produced only ohmic currents that reversed at - 15 mV. After co-expression with Tsha1, a novel outward rectifier current was generated that differed from homomeric Tsha1 by its slower kinetics of activation, its partial current inactivation, and its partial blockade by 5 mM TEA as well as 1 muM DTX. Co-immunoprecipitation studies using anti-Tsha3 antibodies confirmed that Tsha3 tightly bound with Tsha1 in co-infected Sf21 cells. As revealed from GFP- and DsRed-labeling studies, the pattern of distribution of Tsha1 was profoundly altered after co-infection with Tsha3 subunits. (C) 2004 Elsevier B.V. All rights reserved.
ISSN: 0169328X
DOI: 10.1016/j.molbrainres.2004.02.016

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