Secretion of immunoglobulin M assembly intermediates in the presence of reducing agents.
There are several demonstrations that misfolded or unassembled proteins are not transported along the secretory pathway, but are retained intracellularly, generally in the endoplasmic reticulum. For instance, B lymphocytes synthesize but do not secrete IgM, and only the polymeric form of IgM is secreted by plasma cells. The C-terminal cysteine of the mu heavy chain of secreted IgM (residue 575) is involved in the intracellular retention of unpolymerized IgM subunits. Here we report that the addition of reducing agents to the culture medium, at concentrations which do not affect cell viability, terminal glycosylation, or retention of proteins in the endoplasmic reticulum through the KDEL mechanism, induces secretion of IgM assembly intermediates by both B and plasma cells. Free joining (J) chains, which are not normally secreted by plasma cells unless as part of IgM or IgA, are also secreted in the presence of reducing agents. We propose a role for free thiol groups in preventing the unhindered transport of proteins through the secretory pathway. Under the scheme, assembly intermediates interact through their thiol groups between themselves and/or with unknown proteins of the endoplasmic reticulum. Such interactions may be prevented by altering the intracellular redox potential or by site-directed mutagenesis of the relevant cysteine residue(s).
Alberini
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CM
Istituto di Chimica, Facoltà di Medicina, Università degli Studi di Brescia, Italy.
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Milstein
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Sitia
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D000818:Animals; D003545:Cysteine / Q000502:physiology; D006031:Glycosylation; D007074:Immunoglobulin G / Q000378:metabolism; D007144:Immunoglobulin J-Chains / Q000502:physiology; D007075:Immunoglobulin M / Q000378:metabolism* / Q000648:ultrastructure; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008623:Mercaptoethanol / Q000494:pharmacology; D051379:Mice; D010084:Oxidation-Reduction; D010950:Plasma Cells / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D013997:Time Factors; D014407:Tumor Cells, Cultured
D007074:Immunoglobulin G; D007144:Immunoglobulin J-Chains; D007075:Immunoglobulin M; D046911:Macromolecular Substances; D008623:Mercaptoethanol; D003545:Cysteine