title: Rapid degradation of an unassembled immunoglobulin light chain is mediated by a serine protease and occurs in a pre-Golgi compartment.
abstract: The immunoglobulin kappa light chain produced by the CH12 lymphoma is unusual because it is not secreted when expressed in the absence of a heavy chain. Instead, it undergoes rapid intracellular degradation. This degradation is selective, as another light chain expressed in the same cell is not degraded. It is also a property of the CH12 kappa chain itself, since it is degraded rapidly when expressed either in another myeloma cell or in COS-1 fibroblasts. When provided a heavy chain, this kappa chain assembles into IgM and is then protected from proteolysis. The degradation of kappa requires ATP, is sensitive to reduced temperature and to the thiol reagent diamide. Of all the proteolytic inhibitors tested, 3,4-dichloroisocoumarin, L-1-tosylamido-2-phenylethyl chloromethyl ketone, and to a lesser extent 1-chloro-3-tosylamido-7-amino-2-heptanone, inhibit kappa degradation, suggesting the involvement of a serine protease. The degradation of kappa does not require transport to the Golgi complex, nor is it sensitive to a variety of lysosomotropic agents. Both immunofluorescence and the observed association with the endoplasmic reticulum (ER) stress proteins GRP78/BiP and GRP94 indicate that the kappa chain is localized mostly in the ER. When a point mutation which blocks transport to the Golgi complex is introduced into this kappa chain, the association with the stress proteins is enhanced but the rate of degradation is not significantly decreased. We conclude that the CH12 kappa chain is a particularly good substrate for an ER degradation machinery, and that its sensitivity to the protease(s) is governed by its state of assembly. This ER degradation provides a possible quality control mechanism during the differentiation of B lymphocytes.
authors.affiliation: Department of Immunology, Duke University Medical Center, Durham, North Carolina 27710.
authors.forename: A M
authors.initials: AM
authors.lastname: Gardner
authors.forename: S
authors.initials: S
authors.lastname: Aviel
authors.forename: Y
authors.initials: Y
authors.lastname: Argon
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000968:Antimycin A / Q000494:pharmacology; D002352:Carrier Proteins / Q000032:analysis; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D000091342:Endoplasmic Reticulum Chaperone BiP; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000032:analysis; D007145:Immunoglobulin kappa-Chains / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008223:Lymphoma / Q000276:immunology*; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008815:Mice, Inbred Strains; D018832:Molecular Chaperones; D009101:Multiple Myeloma; D017354:Point Mutation; D011480:Protease Inhibitors / Q000494:pharmacology*; D012697:Serine Endopeptidases / Q000378:metabolism*; D013347:Subcellular Fractions / Q000201:enzymology; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D002352:Carrier Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D007145:Immunoglobulin kappa-Chains; D008565:Membrane Proteins; D018832:Molecular Chaperones; D011480:Protease Inhibitors; C018551:glucose-regulated proteins; D000968:Antimycin A; D000255:Adenosine Triphosphate; D012697:Serine Endopeptidases