Secretion of a homodimeric V alpha C kappa T-cell receptor-immunoglobulin chimeric protein.

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TitleSecretion of a homodimeric V alpha C kappa T-cell receptor-immunoglobulin chimeric protein.
Publication TypeJournal Article
Year of Publication1989
AuthorsMariuzza, RA, Winter, G
JournalJ Biol Chem
Volume264
Issue13
Pagination7310-6
Date Published1989 May 5
ISSN0021-9258
KeywordsAmino Acid Sequence, Animals, Base Sequence, DNA, Genes, Immunoglobulin, Glycosylation, Immunoglobulin kappa-Chains, Mice, Myeloma Proteins, Plasmids, Protein Processing, Post-Translational, Receptors, Antigen, T-Cell, Receptors, Antigen, T-Cell, alpha-beta, Recombinant Proteins, Transfection, Tumor Cells, Cultured
Abstract

Transfection into lymphoid cells of a chimeric T-cell receptor-immunoglobulin gene has been used to generate a secreted water-soluble form of the variable (V) domain of a human T-cell receptor alpha chain for use in structural (i.e. x-ray crystallographic) studies. The chimeric protein consists of the V alpha region of the T-cell receptor of a diphtheria toxoid-specific human T-cell clone fused to a human immunoglobulin kappa light chain constant (C) region. It is efficiently secreted by myeloma cells as a noncovalent homodimer of 65-kDa molecular mass in the absence of either immunoglobulin heavy or light chain. The V alpha C kappa protein is extensively glycosylated, and its secretion is glycosylation-dependent. Chimeric genes containing the V beta region of this particular T-cell receptor linked to immunoglobulin C kappa or C gamma 2 regions are expressed intracellularly, but the products, although glycosylated, are not secreted, nor do they assemble with the V alpha C kappa protein. This suggests that the chimeric beta chain-immunoglobulin proteins are incorrectly folded and/or processed due either to the design of the gene fusions themselves or to the absence of vital T-cell-specific accessory molecules in the myeloma host.

Alternate JournalJ. Biol. Chem.
PubMed ID2523393