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PubMed 18502196


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Title: Functional stabilization of Kv1.5 protein by Hsp70 in mammalian cell lines.

Authors: Yutaka Hirota, Yasutaka Kurata, Masaru Kato, Tomomi Notsu, Shunya Koshida, Toshiaki Inoue, Yasushi Kawata, Junichiro Miake, Udin Bahrudin, Peili Li, Yoshiko Hoshikawa, Yasutaka Yamamoto, Osamu Igawa, Yasuaki Shirayoshi, Akira Nakai, Haruaki Ninomiya, Katsumi Higaki, Masayasu Hiraoka, Ichiro Hisatome

Journal, date & volume: Biochem. Biophys. Res. Commun., 2008 Aug 1 , 372, 469-74

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/18502196


Abstract

The aim of this study was to elucidate the mechanisms for regulations of cardiac Kv1.5 channel expression. We particularly focused on the role of heat shock proteins (Hsps). We tested the effects of Hsps on the stability of Kv1.5 channels using biochemical and electrophysiological techniques: co-expression of Kv1.5 and Hsp family proteins in mammalian cell lines, followed by Western blotting, immunoprecipitation, pulse-chase analysis, immunofluorescence and whole-cell patch clamp. Hsp70 and heat shock factor 1 increased the expression of Kv1.5 protein in HeLa and COS7 cells, whereas either Hsp40, 27 or 90 did not. Hsp70 prolonged the half-life of Kv1.5 protein. Hsp70 was co-immunoprecipitated and co-localized with Kv1.5-FLAG. Hsp70 significantly increased the immunoreactivity of Kv1.5 in the endoplasmic reticulum, Golgi apparatus and on the cell membrane. Hsp70 enhanced Kv1.5 current of transfected cells, which was abolished by pretreatment with brefeldin A or colchicine. Thus, Hsp70, but not other Hsps, stabilizes functional Kv1.5 protein.