T cell receptor-extracellular constant regions as hetero-cross-linkers for immunoglobulin variable regions.
The T cell receptor alpha and beta chains are covalently linked via a disulfide bond in their extracellular constant regions. To use these domains as specific hetero-cross-linkers of two different polypeptides, we created genetic constructs encoding a chimeric antibody Fab fragment in which mouse immunoglobulin constant regions from a phosphorylcholine-specific antibody were substituted for human alpha beta-T cell receptor (TCR) extracellular constant regions (for solubilization, the transmembrane- and cytoplasmic-regions of the receptor were deleted). These constructs, i.e., chimeric heavy (VHC beta C kappa) and light (VLC alpha) chains, were cotransfected into murine SP2/0 myeloma cells for expression. Cells transfected with the genes expressed mRNAs for chimeric heavy and light chains. Without CD3 molecules, the two chimeric chains specifically associated via a disulfide bond to form a chimeric Fab fragment in the cells. These data indicate that the TCR C alpha- and C beta-regions might be used as potent specific hetero-cross-linkers for protein engineering.
Seimiya
H
H
Institute of Applied Microbiology, University of Tokyo.
Naito
M
M
Mashima
T
T
Yasui
H
H
Kuwana
Y
Y
Kurosawa
Y
Y
Tsuruo
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D004247:DNA / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D015202:Protein Engineering; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology
D003432:Cross-Linking Reagents; D007135:Immunoglobulin Variable Region; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011993:Recombinant Fusion Proteins; D004247:DNA