中文摘要：

目的验证Caveolin－1蛋白与HERG钾通道是否存在相互作用，并进一步明确发生相互作用的区域。方法（1）将携带不同Caveolin-1片段的pGADT7载体转染Y187，随后分别与转化有pGBKT7－berg—NT或pGBKT7－berg-CT的AHl09进行酵母双杂交；（2）应用免疫共沉淀技术验证Caveolin-1与HERG之间的相互作用；（3）免疫荧光细胞化学分析：pcDNA3．1－HERG和pcDNA3．0－Caveolin-1质粒共转染HEK293细胞，应用特异性抗体和荧光标记二抗显示Caveolin-1及HERG的亚细胞定位，应用激光共聚焦显微镜观察。结果（1）酵母双杂交发现，Caveolin-1与HERG氨基端相互作用，且其寡聚化结构域是必需片段，而Caveolin-1羧基端则与HERG羧基端相互作用；（2）抗Caveolin-1的抗体能够从心肌裂解物中沉淀HERG通道蛋白；（3）Caveolin-1蛋白和HERG通道共定位于细胞膜上。结论Caveolin-1与心脏HERG钾通道存在相互作用，Caveohn-1的寡聚化结构域是HERG氨基片段的相互作用区域，而HERG羧基片段则与Caveolin-1的羧基端相互作用。

英文摘要：

Objectives To confirm the interaction between Caveolin-1 protein and HERG channels, and to identify the interacting domain. Methods （ 1 ）Y187 transformed with pGADT7 carrying different fragments of Caveolin-1 was mated with AH 109 transformed with pGBKT7-herg-NT or pGBKT7-herg-CT, respectively. （2） To further confirm the interaction, co-immunoprecipitation was performed by specific antibody. （3）To study the co-localization between Caveolin-1 and HERG, the anti-Caveolin-1 antibody and anti-HERG antibody were used to probe the subcellular localization of these two proteins by fluorescence immunocytochemistry. Results （1）The yeast hybrid results showed that the oligomerization domain of Caveolin- 1 was essential for the binding of HERG-NT to Caveolin- 1, and Caveolin- 1-CT binding to HERG-CT. （2）The anti- Caveolin-1 antibody precipitated the HERG channel protein from the rat heart lysates. （3）The co-localization of the Caveolin-1 and HERG occurred mostly in the membrane surface compartment, where the majority of HERG presented. Conclusions HERG channels interact with Caveolin-1, while the oligomerization domain of Caveolin-1 is essential for the binding of HERG-NT to Caveolin-1, and Caveolin-l-CT binding to HERG-CT.