A Rapidly Sedimenting Fraction of Rat Liver Endoplasmic Reticulum

Abstract

Balance-sheet experiments carried out to account for the distribution of endoplasmic reticulum fragments during subcellular fractionation of rat liver showed that a large proportion of these fragments are present in the pellets of low-speed centrifugation. Using glucose-6-phosphatase and RNA as markers we found that approximately 50% of the fragments of endoplasmic reticulum sedimented in the pellet of a 640-g centrifugation, 10% in that of a 6000-g centrifugation and 35% in the pellet of a 105000-g centrifugation. Starvation of the animals before use did not alter this distribution, nor did the use of more vigorous homogenization conditions. We have developed a procedure for removing nuclei and erythrocytes from the material sedimenting at 640g to give a fraction (rapidly sedimenting ER fraction or RS-ER) similar to the standard microsomal preparation. Centrifugation of this RS-ER fraction over 1.3 M sucrose yields subfractions of high and low RNA content analogous to the rough and smooth microsomal fractions. Electron-microscopic studies showed that, whereas the rough microsomal fraction consisted of ribosome-studded vesicles of varying size and content density, the rough RS-ER fraction contained a mixture of mitochondria and double lamellar membranes with ribosomes attached. These double lamellar membranes closely resemble the endoplasmic reticulum of intact rat liver. The double lamellar membranes are frequently observed grouped in stacks and in close association with the mitochondria. The significance of the association between endoplasmic reticulum and mitochondria of the RS-ER fraction and the relation between it and the standard microsomal preparation are discussed.