Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum.

作者：Yukio Fujiki, A L Hubbard, Stanley Fowler, Paul B. Lazarow · 发表于：The Journal of Cell Biology · 年份：1982 · DOI：10.1083/jcb.93.1.97 · 被引用次数：1768 · 研究领域：Adipose Tissue and Metabolism、Lipid metabolism and biosynthesis、Mitochondrial Function and Pathology

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.