细胞内分类和 (Na+,K+) ATPase的极化细胞表面传递,这是MDCK细胞底侧血膜的内源成分
Cell
|August 15, 1986
概括
新合成的-ATPaseα子单元被细胞内分类到Madin Darby犬细胞中的底侧膜. 这个过程发生在插入之前,不需要酸性.
科学领域:
- 细胞生物学 细胞生物学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- - ATPase (Na+,K+-ATPase) 是维持细胞离子梯度的关键蛋白质.
- 了解像Na+,K+-ATPase这样的膜蛋白的合成后分类对于细胞功能至关重要.
- 马丁·达比犬 (MDCK) 细胞是研究上皮细胞极性的一种广泛使用的模型.
研究的目的:
- 研究MDCK细胞中Na+,K+-ATPaseα子单元的细胞内分类机制.
- 确定后合成分类的地点,并确定此过程所需的酸性隔间.
主要方法:
- 使用了在双室系统中培养的Madin Darby犬 (MDCK) 细胞.
- 使用N-azidobenzoyl ouabain (NAB-ouabain) 和抗-ouabain抗体进行脉冲标记实验.
- 监测了新合成的Na+,K+-ATPaseα子单元在顶端和底侧表面的到来.
主要成果:
- 新合成的α子单元与NAB-ouabain结合,并且只有当NAB-ouabain处于底侧腔室时才受到免疫沉.
- 不到10%的合成的Na+,K+-ATPase出现在测试中未检测到的顶端表面.
- 证明了Na+,K+-ATPaseα子单元的细胞内分类到底侧域.
结论:
- 在直接插入基侧血之前,Na+,K+-ATPaseα子单元被细胞内分类.
- 排序过程不需要经过酸性隔间.
- 证实MDCK细胞是研究表皮蛋白贩运和极性的一种合适模型.
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