Monoubiquitination modulating membrane trafficking of ROMK channel

• 作者: Yun-longBAI Xiao-yanZHAO JingXIAO XingYAN Dao-hongLIN Bao-fengYANG
• 来源期刊: 中国药理通讯
• 学科: 医学
• 关键词: 单相调节作用 隔膜 ROMK 药物分析
• 年，卷（期）: 2005,（2）
• 页码范围: 14-15
• 页数: 2页
• 分类号: R917

摘要

AIM To study the membrane trafficking modulation of ROMK1 channel by ubiquitination. METHODS we immunoprecipitated protein with ROMK antibody from rat renal cortex and outer medulla, and apply western blot with ubiquitin antibody to detect that ROMK protein is being ubiquitination under physiological condition in vivo. Furthermore, we used His-antibody and ubiquitin antibodies to recongnize proteins in the cotransfected HEK293 cells with hemagglutinin-ubiquitin and His- tagged ROMK1 constructs, respectively. After all intracellular lysine (Lys) residues of ROMK1 had been individually mutated to arginine (Arg), two -electrode voltage clamp was used to measure the ROMK1 channel activity in Xenopus oocytes. To detect the effect of ubiquitin on ROMK1, we used Western blotting technique to search the difference on total protein expression levelspus oocytes.