Stabilizing effects of eicosapentaenoic acid on Kv1.5 channel protein expressed in mammalian cells
| Autor: | Yasutaka Yamamoto, Yutaka Hirota, Haruaki Ninomiya, Masaaki Soma, Osamu Igawa, Akio Yoshida, Yoshiko Hoshikawa, Ting Y. Kuang, Shunya Koshida, Junichiro Miake, Udin Bahrudin, Goshi Shiota, Yasuaki Shirayoshi, Tomomi Notsu, Shingo Harada, Ichiro Hisatome, Peili Li, Katsumi Higaki, Yasutaka Kurata |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Proteasome Endopeptidase Complex
Patch-Clamp Techniques Docosahexaenoic Acids Recombinant Fusion Proteins Blotting Western Action Potentials Biology Transfection Rats Inbred WKY complex mixtures Kv1.5 Potassium Channel chemistry.chemical_compound symbols.namesake Chlorocebus aethiops Animals Immunoprecipitation natural sciences Channel blocker Pharmacology Dose-Response Relationship Drug Thioctic Acid Protein Stability urogenital system Endoplasmic reticulum Fibroblasts Brefeldin A Golgi apparatus Molecular biology Eicosapentaenoic acid Potassium channel Rats Transport protein Eicosapentaenoic Acid nervous system Biochemistry chemistry Docosahexaenoic acid COS Cells symbols lipids (amino acids peptides and proteins) Peptides Oligopeptides |
| Zdroj: | European Journal of Pharmacology. 604:93-102 |
| ISSN: | 0014-2999 |
| Popis: | We investigated the effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on the stability of Kv1.5 channel protein. The expression and function of Kv1.5 (Kv1.5-FLAG) in transfected African green monkey kidney fibroblast cells as well as rat atrium were estimated by immunoblotting, immunoprecipitation, immunofluorescence and patch-clamp techniques. Both EPA and DHA immediately blocked Kv1.5 channel current in a dose-dependent manner, accompanied by reduction of their phosphorylation. Chronic treatment (for 12 h) with EPA at lower concentrations (0.3-10 muM) increased the level of Kv1.5-FLAG protein as well as Kv1.5 channel current without changes in its gating kinetics, prolonging its half-life; in contrast, both EPA and DHA at higher concentrations (30-100 muM) decreased the expression of Kv1.5-FLAG. EPA at the higher concentrations also decreased mRNA of Kv1.5 and synapse-associated protein 97 expression. EPA at the lower concentrations increased Kv1.5 expression in the endoplasmic reticulum, Golgi apparatus and cell membrane. EPA-induced increase of Kv1.5 channel expression and current was abolished by pretreatment with the protein transport inhibitor brefeldin A or colchicines, and by the Kv1.5 channel blocker 4-aminopyridine. Oral administration of EPA (30 mg/kg) increased the level of endogenous Kv1.5 in rat atria. These results indicate that chronic treatment with EPA at lower concentrations stabilizes Kv1.5 channel protein in the endoplasmic reticulum and Golgi apparatus thereby enhancing the Kv1.5 channel current on the cell membrane. |
| Databáze: | OpenAIRE |
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