CAJ | 학술논문

目的：研究电压依赖性钾离子通道-IKs通道的调节亚基KCNE1胞外肽段(氨基端肽段，简称N段)的功能，探究胞外N段在KCNE1调控IKs通道中的作用。方法：通过分子生物学技术制备KCNE1的N段缺失突变体；利用Western Blot方法了解突变体的蛋白表达情况；以细胞免疫染色方法观察突变体的亚细胞定位情况；以全细胞膜片钳记录技术检测突变体(与KCNQ1共表达)的通道电流特征。结果：成功构建KCNE1胞外N段缺失的突变体KCNE1-ΔN，突变体体外表达正常；与野生型相比：KCNE1-ΔN在细胞内的分布出现蛋白集聚现象；通道的电流密度减小，激活电压相比较出现了明显的左移，即通道的激活速率变快。结论：KCNE1的N段对其蛋白表达无明显影响，但对蛋白的转运起重要作用；N段参与了KCNE1对通道电流大小的调节，以及对通道激活特征的调控。
목적：연구전압의뢰성갑리자통도-IKs통도적조절아기KCNE1포외태단(안기단태단，간칭N단)적공능，탐구포외N단재KCNE1조공IKs통도중적작용。방법：통과분자생물학기술제비KCNE1적N단결실돌변체；이용Western Blot방법료해돌변체적단백표체정황；이세포면역염색방법관찰돌변체적아세포정위정황；이전세포막편겸기록기술검측돌변체(여KCNQ1공표체)적통도전류특정。결과：성공구건KCNE1포외N단결실적돌변체KCNE1-ΔN，돌변체체외표체정상；여야생형상비：KCNE1-ΔN재세포내적분포출현단백집취현상；통도적전류밀도감소，격활전압상비교출현료명현적좌이，즉통도적격활속솔변쾌。결론：KCNE1적N단대기단백표체무명현영향，단대단백적전운기중요작용；N단삼여료KCNE1대통도전류대소적조절，이급대통도격활특정적조공。
Objective: To investigate the function of N region in KCNE1, which is the regulatory subunit of voltage gated potassium channel-IKs, in order to find out the role of N region on KCNE1 modulating channel. Methods: With molecular technologies, we created N-truncated form named KCNE1-ΔN and then analyse its protein expression using Western Blot;immunocytochemistry was employed to detect subcellular localization of the truncation; while whole cell patch-calmp was used to detect the characteristics of KCNE1-ΔN expressed with KCNQ1. Results: KCNE1-ΔN was constructed successfully, and expressed normally in vivo. Comparing to wide type of KCNE1, KCNE1-ΔN showed intracellular aggression;KCNE1-ΔN also showed significant reduction on whole cell current densities and displayed a significant shift of the voltage-dependent curve towards left, which means speeded activation. Conclusions: N region does not have effect on protein expression but playes a significant role on protein trafficking; N region plays a role on KNCE1 modulating IKs channel’s whole cell current densities and channel characteristics on activation.