A single amino acid substitution in the variable region of the light chain specifically blocks immunoglobulin secretion.
Although immunoglobulin light chains are usually secreted in association with heavy chains, free light chains can be secreted by lymphocytes. To identify the structural features of light chains that are essential for their secretion, we mutated a conserved sequence in the variable domain of a lambda I light chain. The effects of the mutations on secretion were assayed by transient expression in COS-1 cells. One mutant (AV60), which replaced Ala-60 with Val, was secreted as efficiently as wild-type lambda I by transfected COS-1 cells. This result was not surprising because secreted lambda II chains contain valine in this position. However, a second lambda I mutant (AV60FS62), which replaced Phe-62 with Ser as well as Ala-60 with Val, was not secreted. This mutant was arrested in the endoplasmic reticulum, as judged by immunofluorescence and by its association with a lumenal endoplasmic reticulum protein, immunoglobulin heavy chain binding protein (BiP). The defect in secretion was not due to gross misfolding of the lambda I chain, since cells cotransfected with AV60FS62 and an immunoglobulin heavy chain gene produced functional antigen-binding antibodies. These assembled IgM molecules were still not secreted. Hence, the replacement of Phe-62 with Ser specifically affects a determinant on the lambda I light chain that is necessary for the intracellular transport of this molecule.
Dul
J L
JL
Department of Microbiology and Immunology, Duke University Medical Center, Durham, NC 27710.
Argon
Y
Y
D000818:Animals; D002460:Cell Line; D005803:Genes, Immunoglobulin; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D051379:Mice; D008961:Models, Structural; D016297:Mutagenesis, Site-Directed; D010649:Phenylalanine; D010954:Plasmacytoma; D011487:Protein Conformation; D015183:Restriction Mapping; D012694:Serine; D014162:Transfection; D014633:Valine
D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D012694:Serine; D010649:Phenylalanine; D014633:Valine