title: In vivo cross-linking of protein disulfide isomerase to immunoglobulins.
abstract: To test the proposed role of protein disulfide isomerase in the synthesis of immunoglobulins (Ig), intact lymphocytes were treated with a thiol-cleavable, bifunctional cross-linking agent and lysed, and the lysates were immunoprecipitated with antibodies to either Ig or enzyme. When the immunoprecipitates were analyzed on polyacrylamide-sodium dodecyl sulfate gels, protein disulfide isomerase was found to be cross-linked to immunoglobulins. The extent of cross-linking was dependent upon the concentration of cross-linker added and the class of Ig. For IgMs and high concentrations of cross-linker, approximately one molecule of Ig was coupled per two molecules of enzyme. For IgGs, the extent of cross-linking was less. Finally, depletion of the intracellularly reduced glutathione by diamide was found to also result in the linkage of protein disulfide isomerase to IgM. These results therefore support the hypothesis that protein disulfide isomerase functions in the in vivo synthesis of immunoglobulins.
authors: Department of Pharmacology, Stanford University School of Medicine, California 94305-5332.
authors: R A
authors: RA
authors: Roth
authors: S B
authors: SB
authors: Pierce
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003958:Diamide / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D006825:Hybridomas / Q000201:enzymology / Q000276:immunology; D007070:Immunoglobulin A / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D007535:Isomerases / Q000378:metabolism*; D019704:Protein Disulfide-Isomerases
chemical_list: D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D007070:Immunoglobulin A; D007075:Immunoglobulin M; D003958:Diamide; D007535:Isomerases; D019704:Protein Disulfide-Isomerases; D005978:Glutathione