Isolation and properties of polyribosomes and fragments of the endoplasmic reticulum from rat liver

Abstract

A centrifugation method for the fractionation of the postmitochondrial fraction from rat-liver homogenates is described. The technique, in which no detergent is used, may be used as a tool to discriminate between two classes of ribosomes. One class is firmly bound to membranes and the other consists either of free polysomes or of ribosomes attached by weaker forces to the membranes of the endoplasmic reticulum. Electron-micrograph studies revealed that the polysomes were not contaminated with bound ribosomes or with membranous fragments. The separated fractions were characterized by their RNA, protein, ribonuclease and phospholipid content. The influence of starvation on the RNA and protein contents of the different fractions was investigated. Labelling of the various centrifugal fractions in vivo revealed no difference in uptake of radioactive amino acid between the two classes of ribosomes. Incorporation of radioactive leucine in vitro and the polyuridylic acid-directed phenylalanine incorporation were similar for both classes of ribosomes.