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

被引：16

作者：

Hirota, Yutaka ^{[2
]}

Kurata, Yasutaka ^{[1
]}

Kato, Masaru ^{[3
]}

Notsu, Tomomi ^{[2
]}

Koshida, Shunya ^{[2
]}

Inoue, Toshiaki ^{[4
]}

Kawatae, Yasushi ^{[5
]}

Miake, Junichiro ^{[3
]}

Bahrudin, Udin ^{[2
]}

Li, Peili ^{[2
]}

Hoshikawa, Yoshiko ^{[2
]}

Yamamoto, Yasutaka ^{[2
]}

Igawa, Osamu ^{[3
]}

Shirayoshi, Yasuaki ^{[2
]}

Nakai, Akira ^{[6
]}

Ninomiya, Haruaki ^{[7
]}

Higaki, Katsumi ^{[8
]}

Hiraoka, Masayasu ^{[9
]}

Hisatome, Ichiro ^{[2
]}

机构：

[1] Kanazawa Med Univ, Dept Physiol, Uchinada, Ishikawa 9200268, Japan

[2] Tottori Univ, Div Regenerat Med & Therapeut, Dept Genet Med & Regenerat Therapeut, Inst Regenerat Med & Biofunct,Grad Sch Med Sci, Yonago, Tottori 683, Japan

[3] Tottori Univ, Fac Med, Dept Cardiovasc Med, Yonago, Tottori 683, Japan

[4] Tottori Univ, Fac Med, Dept Human Genome Sci, Yonago, Tottori 683, Japan

[5] Tottori Univ, Div Prot Funct, Dept Biotechnol, Fac Engn, Tottori 6808552, Japan

[6] Yamaguchi Univ, Sch Med, Dept Biochem & Mol Biol, Ube, Yamaguchi 755, Japan

[7] Tottori Univ, Fac Med, Dept Neurobiol, Yonago, Tottori 683, Japan

[8] Tottori Univ, Ctr Gene Res, Yonago, Tottori 683, Japan

[9] Minist Hlth Labor & Welf, Labor Insurance Appeal Comm, Minato Ku, Tokyo 1050011, Japan

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2008年 / 372卷 / 03期

关键词：

cardiac Kv1.5 channel; heat shock proteins; western blotting; immunoprecipitation; pulse-chase analysis; immunofluorescence; whole-cell patch clamp; COS7; cells;

D O I：

10.1016/j.bbrc.2008.05.068

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 colchincine. Thus, Hsp70, but not other Hsps. stabilizes functional Kv1.5 protein. (C) 2008 Elsevier Inc. All rights reserved.

引用

页码：469 / 474

页数：6

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