The secretory pathways of rat serum glycoproteins and albumin. Localization of newly formed proteins within the endoplasmic reticulum.
These studies compare the secretory pathways of newly formed rat serum glycoproteins and albumin by studying their submicrosomal localization at early times after the beginning of their synthesis and also by determining the submicrosomal site of incorporation of N-acetylglucosamine, mannose, galactose, and leucine into protein. N-acetylglucosamine, mannose, and galactose were only incorporated in vitro into proteins from membrane-attached polysomes and not into proteins from free polysomes. Mannose incorporation occurred in the rough endoplasmic reticulum, was stimulated by puromycin but not by cycloheximide, and 90% of the mannose-labeled protein was bound to the membranes. Galactose incorporation, by contrast, occurred in the smooth microsome fraction and 89% of the radioactive protein was in the cisternae. Albumin was mostly recovered (98%) in the cisternae, with negligible amounts in the membranes. To determine whether the radio-active sugars were being incorporated into serum proteins or into membrane protein, the solubilized in vivo-labeled proteins were treated with specific antisera to rat serum proteins or to albumin. Immunoelectrophoresis of the (14)C-labeled leucine membrane and cisternal proteins showed that the membranes contained radioactive serum glycoprotein but no albumin, while the cisternal fraction contained all of the radioactive albumin and some glycoproteins. The results indicate that newly formed serum glycoproteins remain attached to the membranes of the rough endoplasmic reticulum after they are released from the membrane-attached polysomes, while albumin passes directly into the cisternae.
D000818:Animals; D001345:Autoradiography; D002247:Carbon Isotopes; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System; D002854:Chromatography, Paper; D003513:Cycloheximide / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005690:Galactose / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism; D006023:Glycoproteins / Q000097:blood / Q000378:metabolism*; D007122:Immunoelectrophoresis; D007930:Leucine / Q000378:metabolism; D008358:Mannose / Q000378:metabolism; D008861:Microsomes / Q000187:drug effects; D008862:Microsomes, Liver / Q000378:metabolism*; D008954:Models, Biological; D014176:Protein Biosynthesis; D011691:Puromycin / Q000494:pharmacology; D051381:Rats; D012270:Ribosomes / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism*; D014316:Tritium
D002247:Carbon Isotopes; D006023:Glycoproteins; D012709:Serum Albumin; D014316:Tritium; D011691:Puromycin; D003513:Cycloheximide; D007930:Leucine; D005944:Glucosamine; D008358:Mannose; D005690:Galactose