Translocation of nascent secretory proteins across membranes can occur late in translation.
Signal recognition particle (SRP) causes an arrest in the translation of nascent secretory proteins in a wheat germ cell-free system. In order to examine at what point during the synthesis of a secretory protein its translocation across the endoplasmic reticulum (ER) membrane can occur, SRP was used to arrest nascent chain elongation at various times during a synchronous translation, thus allowing the generation of nascent chains of increasing length. It was found that SRP can still bring about an arrest as late as when an average of two-thirds of nascent IgG light chain was completed. Rough microsomes were added to translations blocked with SRP to determine if such relatively long nascent chains could still be translocated across the membrane. It was found that nascent chains which had been arrested by SRP, regardless of their length, could be translocated into rough microsomes. In the case of IgG light chain, translocation levels of 50% were still observed with nascent chains corresponding to as much as 70-75% of the intact preprotein. Similar results were observed for the nascent bovine prolactin precursor. These results demonstrate that the synthesis of secretory proteins can be uncoupled from their translocation, and that fairly large nascent chains are capable of crossing the membrane of the ER post-translationally.
D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000378:metabolism; D007074:Immunoglobulin G / Q000235:genetics*; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007700:Kinetics; D051379:Mice; D008861:Microsomes / Q000378:metabolism; D010179:Pancreas / Q000378:metabolism; D010441:Peptide Chain Elongation, Translational; D010944:Plants / Q000378:metabolism; D010954:Plasmacytoma / Q000276:immunology; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D014908:Triticum / Q000378:metabolism
D007074:Immunoglobulin G; D007147:Immunoglobulin Light Chains; D012333:RNA, Messenger