The association in vitro of polyribosomes with ribonuclease-treated derivatives of hepatic rough endoplasmic reticulum. Characteristics of the membrane binding sites and factors influencing association.
1. Pancreatic ribonuclease in dilute EDTA has been shown to condition rough-microsomal membranes from adult rat liver to accept exogenously added rat liver polyribosomes in vitro at 0-4 degrees C. Treated smooth membranes would not significantly interact with polyribosomes. 2. The conditioning process decreased the membrane RNA content and removed polyribosomes from vesicle surfaces as viewed electron-microscopically. 3. Binding to these conditioned membranes was shown to be uninfluenced by changes of temperature (0-37 degrees C) and pH (6.9-7.8) or the presence of cell sap, but was inhibited by increasing the concentration of potassium chloride. 4. Possession of a polyribosome-binding capacity by conditioned rough membranes was not dependent on adventitious materials that could be dislodged by high ionic strengths. 5. Trypsin treatment under mild conditions destroyed the binding capacity of ribonuclease-conditioned rough membranes. 6. A 2-10S residual RNA was recovered from ribonuclease-conditioned membranes, but its partial removal had no effect on the capacity of membranes to accept polyribosomes. However, some role for this residual RNA in attaching polyribosomes could not be discounted. 7. Evidence is considered that polyribosome-binding sites are intrinsic features of conditioned membranes isolated from rough-microsomal fractions, and that long-range ionic bonding is a primary factor in polyribosome interaction with these binding sites.
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002247:Carbon Isotopes; D002499:Centrifugation, Density Gradient; D004721:Endoplasmic Reticulum / Q000032:analysis / Q000187:drug effects*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008297:Male; D008854:Microscopy, Electron; D008862:Microsomes, Liver / Q000187:drug effects*; D010759:Phosphorus Isotopes; D011189:Potassium Chloride / Q000494:pharmacology; D012313:RNA / Q000032:analysis; D012335:RNA, Ribosomal; D051381:Rats; D012260:Ribonucleases / Q000494:pharmacology*; D012270:Ribosomes; D013696:Temperature; D014357:Trypsin / Q000494:pharmacology
D000596:Amino Acids; D002247:Carbon Isotopes; D010759:Phosphorus Isotopes; D012335:RNA, Ribosomal; D012313:RNA; D011189:Potassium Chloride; D012260:Ribonucleases; D014357:Trypsin