The selectivity and stoicheiometry of membrane binding sites for polyribosomes, ribosomes and ribosomal subunits in vitro.
Differences in the binding sites for polyribosomes, template-depleted ribosomes and large ribosomal subunits were found in microsomal derivatives of the rough endoplasmic reticulum. 1. The stoicheiometry of polyribosome and ribosome interaction in vitro with membranes was shown to be influenced by the relative concentration of interactants and the duration of their mixing. Large ribosomal subunits required a more prolonged mixing schedule to achieve saturation of membranes than did polyribosomes. 2. By using a procedure which minimized the effects on binidng by the stoicheiometric variables, competition between populations of polyribosomes, ribosomes and subunits for membrane sites showed that subunits, and to a lesser extent ribosomes, failed to block polyribosome attachment. 3. Polyribosomes isolated from liver, kidney and hepatoma 5123C entirely bound to a common membrane site, but some polyribosomes from myeloma MOPC-21 bound to other sites, perhaps influenced by their unique nascent proteins. 4. Subunit-binding sites appear on rough membranes only after endogenous polyribosomes have been removed, but no evidence that resulting changes in surface constituents are responsible was found. Large-subunit binding was largely abolished by lowering MgC12 concentration of 0.1 mM, whereas under the same conditions polyribosome binding was undiminished. 5. The large-subunit site appears to be distinct from the polyribosome site not only in the restriction of its affinity for particles but also spatially, to the extent that bound subunits do not hinder access of polyribosomes to their sites.
D000818:Animals; D001665:Binding Sites; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002458:Cell Fractionation; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000378:metabolism; D066298:In Vitro Techniques; D007668:Kidney / Q000648:ultrastructure; D008099:Liver / Q000648:ultrastructure; D008113:Liver Neoplasms; D008274:Magnesium; D008297:Male; D008566:Membranes / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism*; D009101:Multiple Myeloma / Q000378:metabolism; D011132:Polyribosomes / Q000032:analysis / Q000378:metabolism; D051381:Rats; D012270:Ribosomes / Q000378:metabolism*
D008274:Magnesium