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Engineering Cell-permeable Protein
Published on: December 28, 2009
在稳定表达囊性纤维化基因的CHO细胞中,酸化调节的Cl通道稳定表达囊性纤维化基因
J A Tabcharani1, X B Chang, J R Riordan
1Department of Physiology, McGill University, Montreal, Québec, Canada.
Nature
|August 15, 1991
概括
囊性纤维化基因在非上皮细胞中表达化通道,由循环AMP激活,并通过酸化调节. 这项研究证明了囊性纤维化跨膜调节道的直接酸化控制.
科学领域:
- 分子生物学分子生物学
- 细胞生理学 细胞生理学
- 离子通道功能的功能
背景情况:
- 已知囊性纤维化基因产物,囊性纤维化转膜调节器 (CFTR) 调节化通道.
- 循环AMP (cAMP) 刺激的化物导电性已在由低导电性Cl-通道介导的上皮细胞中观察到.
- 在非上皮细胞中缺乏对该通道酸化控制的直接证明.
研究的目的:
- 研究通过非上皮细胞系统的酸化对囊性纤维化跨膜调节器 (CFTR) 化道的直接调节.
- 描述CFTR介导的化物导电性的功能性质和调节机制.
主要方法:
- 具有囊性纤维化基因的中国仓鼠卵巢 (CHO) 细胞的稳定转移.
- 电子生理学记录以测量化物通道活性.
- 通过膜穿透的cAMP类似物和纯化蛋白激酶/酸酶的应用,以研究通道调节.
主要成果:
- 在CHO细胞中,囊性纤维化基因的稳定表达导致了低导电率的欧姆Cl-通道的出现.
- 这种通道活性是由通过膜穿透的cAMP类似物应用于细胞表面 (在细胞上) 刺激的.
- 当应用到膜的细胞内侧 (细胞外) 时,通道活性由蛋白激酶A和C (酸化) 和性酸酶 (脱酸化) 调节.
结论:
- 囊性纤维化转膜调节器作为化物通道,由酸化和脱酸化事件直接调节.
- 收的激酶调节通路在控制CFTR通道活性方面表现出协同作用.
- 这些发现为CFTR作为受调节的离子通道的作用提供了有力的证据,并突出了复杂的调节机制.
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