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Covalent assembly of a soluble T cell receptor-peptide-major histocompatibility class I complex.
- Source :
-
Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 1996 Jul 09; Vol. 93 (14), pp. 7184-9. - Publication Year :
- 1996
-
Abstract
- We used stepwise photochemical cross-linking for specifically assembling soluble and covalent complexes made of a T-cell antigen receptor (TCR) and a class I molecule of the major histocompatibility complex (MHC) bound to an antigenic peptide. For that purpose, we have produced in myeloma cells a single-chain Fv construct of a TCR specific for a photoreactive H-2Kd-peptide complex. Photochemical cross-linking of this TCR single-chain Fv with a soluble form of the photoreactive H-2Kd-peptide ligand resulted in the formation of a ternary covalent complex. We have characterized the soluble ternary complex and showed that it reacted with antibodies specific for epitopes located either on the native TCR or on the Kd molecules. By preventing the fast dissociation kinetics observed with most T cell receptors, this approach provides a means of preparing soluble TCR-peptide-MHC complexes on large-scale levels.
- Subjects :
- Affinity Labels
Amino Acid Sequence
Animals
Base Sequence
Binding, Competitive
Cell Line
Cross-Linking Reagents
DNA Primers
H-2 Antigens isolation & purification
H-2 Antigens metabolism
Hybridomas
Kinetics
Major Histocompatibility Complex
Mice
Molecular Sequence Data
Polymerase Chain Reaction
Receptors, Antigen, T-Cell isolation & purification
Receptors, Antigen, T-Cell metabolism
Receptors, Antigen, T-Cell, alpha-beta isolation & purification
Receptors, Antigen, T-Cell, alpha-beta metabolism
Recombinant Proteins immunology
Recombinant Proteins isolation & purification
Recombinant Proteins metabolism
Tumor Cells, Cultured
H-2 Antigens immunology
Receptors, Antigen, T-Cell immunology
Receptors, Antigen, T-Cell, alpha-beta immunology
T-Lymphocytes, Cytotoxic immunology
Subjects
Details
- Language :
- English
- ISSN :
- 0027-8424
- Volume :
- 93
- Issue :
- 14
- Database :
- MEDLINE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Publication Type :
- Academic Journal
- Accession number :
- 8692966
- Full Text :
- https://doi.org/10.1073/pnas.93.14.7184