NDLI: Functional three-domain single-chain T-cell receptors.

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Functional three-domain single-chain T-cell receptors.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Hafler, D. A. Chung, S. Friedman, S. M. Wucherpfennig, K. W. Strominger, J. L.
Description	Author Affiliation: Chung S ( Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.);
Abstract	T-cell receptors (TCRs) are membrane anchored heterodimers structurally related to antibody molecules. Single-chain antibodies can be engineered by linking the two variable domains, which fold properly by themselves. However, proper assembly of the variable domains of a human TCR (V alpha and V beta) that recognize the HLA-DR2b/myelin basic protein-(85-99) peptide complex was critically dependent on the addition of a third domain, the constant region of the TCR beta chain (C beta), to the single-chain construct. Single-chain molecules with the three-domain design, but not those with the two-domain design, expressed in a eukaryotic cell as chimeric molecules linked either to glycosyl phosphatidylinositol or to the transmembrane/cytoplasmic domains of the CD3 zeta chain were recognized by a conformation-sensitive monoclonal antibody. The chimeric three-domain single-chain TCR linked to CD3 zeta chain signaled in response to both the specific HLA-DR/peptide and the HLA-DR/superantigen staphylococcal enterotoxin B complexes. Thus, by using this three-domain design, functional single-chain TCR molecules were expressed with high efficiency. The lipid-linked single-chain TCR was solubilized by enzymatic cleavage and purified by affinity chromatography. The apparent requirement of the constant domain for cooperative folding of the two TCR variable domains may reflect significant structural differences between TCR and antibody molecules.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	26
Volume Number	91
Language	English
Publisher	National Academy of Sciences
Publisher Date	1994-01-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Receptors, Antigen, T-Cell, Alpha-beta Chemistry Amino Acid Sequence Cloning, Molecular DNA Primers Epitope Mapping Molecular Sequence Data Immunology Recombinant Proteins Serotonin Secretion Signal Transduction Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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