The specificity and stability of the triton-extracted cytoskeletal framework of gerbil fibroma cells.
The specificity and stability of the triton-extracted cytoskeletal framework of gerbil fibroma cells.
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Triton 提取的沙鼠纤维瘤细胞细胞骨架框架的特异性和稳定性。
DOI：
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发表时间：
1985
影响因子：
4
通讯作者：
Alice B. FULTONf
中科院分区：
文献类型：
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作者：
M. Gilbert;Alice B. FULTONf
沙鼠纤维瘤细胞的细胞网和地形可以通过使用Triton X-100的温和提取程序来保存。我们通过测量提取率及其对外源蛋白的敏感性来确定这些细胞骨架框架的稳定性和特异性。使用两种缓冲液，其模拟细胞内和细胞外离子环境。用这两种缓冲液，提取几乎是在5分钟完成。这种模式的提取被认为是在5和9天大的文化。当在平板接种后第二天检查三种不同的细胞稀释液时，观察到相同的提取模式。因此，提取率在很大程度上是独立的离子组成，年龄在文化，或细胞密度的微小变化。通过与两种不同的外源蛋白（牛血清白蛋白或卵清蛋白）的竞争来确定如此产生的细胞骨架框架的特异性，所述外源蛋白没有从细胞骨架框架中去除任何额外的材料，即使在提取缓冲液中具有超过10%的外源蛋白。这种提取模式并不是沙鼠纤维瘤细胞所独有的。在一系列细胞中观察到类似的提取模式：小鼠3T3细胞、3T6细胞和SVPY 3T3细胞。这些实验表明，在适当条件下通过Triton提取产生的细胞骨架框架在提取10分钟或更长时间后是稳定的，并且结构是特异性的，因为它们不被外源蛋白的存在破坏。
Cellular meshworks and topography of gerbil fibroma cells can be preserved by gentle extraction procedures using Triton X-100. We determined the stability and specificity of these cytoskeletal frameworks by measuring extraction rate and its sensitivity to exogenous protein. Two buffers were used, which mimicked the intracellular and extracellular ionic environments. With both buffers, extraction was nearly complete at 5 min. This pattern of extraction was seen both in 5- and 9-day-old cultures. The same pattern of extraction was seen when three different dilutions of cells were examined the second day after plating. Thus, extraction rate was largely independent of minor variations in ionic composition, age in culture, or cell density. Specificity of the cytoskeletal frameworks so produced was determined by competition with two different exogenous proteins (bovine serum albumin or ovalbumin), which did not remove any additional material from the cytoskeletal frameworks, even with over 10% exogenous protein in the extraction buffer. This pattern of extraction is not unique to gerbil fibroma cells. A similar pattern of extraction was seen for a series of cells: mouse 3T3 cells, 3T6 cells and SVPY 3T3 cells. These experiments indicate that the cytoskeletal framework produced by Triton extraction under appropriate conditions is stable after extraction for a period of 10 min or longer, and that the structures are specific, in that they are not disrupted by the presence of exogenous proteins.