通过碳酸钠处理分离细胞内膜：应用于内质网
Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum.
作者信息
Fujiki Y, Hubbard A L, Fowler S, Lazarow P B
出版信息
J Cell Biol. 1982 Apr;93(1):97-102. doi: 10.1083/jcb.93.1.97.
Abstract
A rapid and simple method for the isolation of membranes from subcellular organelles is described. The procedure consists of diluting the organelles in ice-cold 100 mM Na2CO3 followed by centrifugation to pellet the membranes. Closed vesicles are converted to open membrane sheets, and content proteins and peripheral membrane proteins are released in soluble form. Here we document the method by applying it to various subfractions of a rat liver microsomal fraction, prepared by continuous density gradient centrifugation according to Beaufay et al. (1974, J. Cell Biol. 61:213-231). The results confirm and extend those of previous investigators on the distribution of enzymes and proteins among the membranes of the smooth and rough endoplasmic reticulum. In the accompanying paper (1982, J. Cell Biol. 93:103-110) the procedure is applied to peroxisomes and mitochondria.
摘要
本文描述了一种从亚细胞器中分离膜的快速简便方法。该方法包括将细胞器在冰冷的100 mM Na2CO3中稀释，然后离心使膜沉淀。封闭的囊泡转化为开放的膜片，内容物蛋白和外周膜蛋白以可溶形式释放。在此，我们通过将该方法应用于大鼠肝微粒体部分的各种亚组分来记录该方法，这些亚组分是根据Beaufay等人（1974年，《细胞生物学杂志》61:213 - 231）的方法通过连续密度梯度离心制备的。结果证实并扩展了先前研究者关于酶和蛋白质在光滑和粗糙内质网的膜之间分布的研究结果。在随附的论文（1982年，《细胞生物学杂志》93:103 - 110）中，该方法应用于过氧化物酶体和线粒体。
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