Receptor-mediated ribosome binding to liposomes depends on lipid composition.
Ribosome binding to the endoplasmic reticulum has been traditionally studied using an in vitro assay in which potential ribosome receptors have been purified, incorporated into synthetic liposomes, and tested for activity. One such receptor (180 kDa; "p180") has been shown to bind ribosomes with high affinity in such a system when purified to homogeneity. This result has been challenged by data generated in other laboratories, and as a result, doubt has lingered as to the authenticity of p180 as a ribosome receptor. The contribution of the major difference between these studies, the lipid composition of the liposomes used in the in vitro assays, was assessed when identical fractions of rough endoplasmic reticulum-specific membrane proteins were incorporated into liposomes composed of only phosphatidylcholine (as used in other laboratories), a 50:50 mix of phosphatidylcholine and phosphatidylserine (as used in our original studies), or lipids derived from canine pancreatic microsomes (as a physiologically relevant control). The presence of PS was found to be crucial for the incorporation into and ribosome binding activity of p180 in liposomes. These observations are compatible with published studies on the importance of acidic phospholipids in ribosome binding to intact microsomes and reconcile the apparently conflicting in vitro results surrounding the assignment of p180 as a ribosome receptor.
Savitz
A J
AJ
Department of Biological Chemistry and the Molecular Biology Institute, UCLA School of Medicine, Los Angeles, California 90024-1737, USA.
Meyer
D I
DI
D000818:Animals; D004285:Dogs; D050356:Lipid Metabolism; D008055:Lipids / Q000032:analysis; D008081:Liposomes / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D008055:Lipids; D008081:Liposomes; D018160:Receptors, Cytoplasmic and Nuclear; D012269:Ribosomal Proteins