Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum.
Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum.
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DOI：
10.1083/jcb.93.1.97
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发表时间：
1982-04
期刊：
影响因子：
--
通讯作者：
Lazarow PB
中科院分区：
文献类型：
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作者：
Fujiki Y;Hubbard AL;Fowler S;Lazarow PB
本文描述了一种从亚细胞器中分离膜的快速且简便的方法。该操作包括在冰冷的100 mM碳酸钠中稀释细胞器，然后离心使膜形成沉淀。封闭的囊泡转变为开放的膜片，内容物蛋白和外周膜蛋白以可溶形式释放出来。在此，我们通过将该方法应用于大鼠肝脏微粒体组分的各种亚组分来记录此方法，该微粒体组分是按照博费（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。这些结果证实并扩展了先前研究人员关于酶和蛋白质在光面内质网和粗面内质网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，该方法被应用于过氧化物酶体和线粒体。
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.