A Polysome-Membrane Binding System from Rat Liver

I. Basic Characterization of the Binding System

Abstract

A simple reaction system was developed to examine the binding of polysomes to membranes of the endoplasmic reticulum and to investigate the fate of ribosomes and nascent chains during protein synthesis in vitro. The system consisted of Sephadex G-25 treated post-mitochondrial fraction prepared from rat liver (Sephadex-PM) as a source of membranes, and radioactive free polysomes prepared from another rat liver. The following results were obtained.
1. Nascent chains on free polysomes labeled in vivo were transferred to membranes in vitro. The process required protein synthesis.
2. This reaction occurred in two steps: a) Binding of the free polysomes to membranes in the absence of protein synthesis. b) Release of ribosomes, leaving nascent chains on the membranes, requiring protein synthesis.
3. A portion of the ribosomes found on membranes in vivo (membrane-bound ribo-somes) was also released from the membranes during incubation in vitro, leaving their nascent chains on the membranes.
The significance of the transfer of nascent chains from free polysomes to membranes in vitro is discussed in the light of known polysome-membrane interaction in vivo.