Co-translational modification of nascent immunoglobulin heavy and light chains.
We have investigated the in vivo co-translational covalent modification of nascent immunoglobulin heavy and light chains. Nascent polypeptides were separated from completed polypeptides by ion-exchange chromatography of solubilized ribosomes on QAE-Sephadex. First, we have demonstrated that MPC 11 nascent heavy chains are quantitatively glycosylated very soon after the asparaginyl acceptor site passes through the membrane into the cisterna of the rough endoplasmic reticulum. Nonglycosylated completed heavy chains of various classes cannot be glycosylated after release from the ribosome, due either to rapid intramolecular folding and/or intermolecular assembly, which cause the acceptor site to become unavailable for the glycosylation enzyme. Second, we have shown that the formation of the correct intrachain disulfide loop within the first light chain domain occurs rapidly and quantitatively as soon as the appropriate cysteine residues of the nascent light chain pass through the membrane into the cisterna of the endoplasmic reticulum. The intrachain disulfide loop in the second or constant region domain of the light chain is not formed on nascent chains, because one of the cysteine residues involved in this disulfide bond does not pass through the endoplasmic reticulum membrane prior to chain completion and release from the ribosome. Third, we have demonstrated that some of the initial covalent assembly (formation of interchain disulfide bonds) occurs on nascent heavy chains prior to their release from the ribosome. The results are consistent with the pathway of covalent assembly of the cell line, in that completed light chains are assembled onto nascent heavy chains in MPC 11 cells (IgG2b), where a heavy-light half molecule is the major initial covalent intermediate; and completed heavy chains are assembled onto nascent heavy chains in MOPC 21 cells (IgG1), where a heavy chain dimer is the major initial disulfide linked intermediate.
D000818:Animals; D002460:Cell Line; D004220:Disulfides / Q000378:metabolism; D005944:Glucosamine / Q000494:pharmacology; D006027:Glycosides / Q000378:metabolism; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis*; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis*; D051379:Mice; D009374:Neoplasms, Experimental / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D010954:Plasmacytoma / Q000378:metabolism*; D014176:Protein Biosynthesis; D011487:Protein Conformation; D014166:Transferases / Q000378:metabolism
D004220:Disulfides; D006027:Glycosides; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D009844:Oligosaccharides; D014166:Transferases; D005944:Glucosamine