Photoaffinity labeling of the T cell receptor on cloned cytotoxic T lymphocytes by covalent photoreactive ligand.

Détails

ID Serval
serval:BIB_60B6C9BB6356
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Photoaffinity labeling of the T cell receptor on cloned cytotoxic T lymphocytes by covalent photoreactive ligand.
Périodique
The Journal of biological chemistry
Auteur(s)
Luescher I.F., Cerottini J.C., Romero P.
ISSN
0021-9258
Statut éditorial
Publié
Date de publication
1994
Peer-reviewed
Oui
Volume
269
Numéro
8
Pages
5574-5582
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Résumé
The interaction of the T cell antigen receptor with a photoreactive antigenic peptide derivative bound covalently to the H-2Kd (Kd) molecule was studied by photoaffinity labeling on cloned, CD8 positive cytotoxic T lymphocytes. The Kd-restricted Plasmodium berghei circumsporozoite peptide 253-260 (YIPS-AEKI) was conjugated with iodo-4-azidosalicylic acid at the N terminus and with 4-azidobenzoic acid at the T cell receptor residue Lys-259. Cell-associated or soluble Kd molecules were photoaffinity-labeled with the peptide derivative by selective photoactivation of the N-terminal photoreactive group. Incubation of cell-associated or soluble covalent Kd-peptide derivative complexes (ligands) with cytotoxic T lymphocytes that recognized this peptide derivative and activation of the orthogonal photoreactive group resulted in specific photoaffinity labeling of the T cell receptor. The labeling was inhibitable by an anti-Kd antibody and was absent on Kd-restricted cytotoxic T lymphocytes of different specificity. The binding of the soluble ligand reached a maximum after 2-4 min at 37 degrees C, after 30 min at 18 degrees C, and after 3 h at 4 degrees C. In contrast, binding of the cell-associated ligand reached a transient maxima after 50 and 110 min at 37 and 18 degrees C, respectively. The degree of binding at 37 degrees C was approximately 30% lower than that at 18 degrees C. No binding took place at 4 degrees C. Inhibition studies with antibodies and drugs indicated that the binding of the cell-associated, but not the soluble ligand, was highly dependent on T cell-target cell conjugate formation, whereas the binding of the soluble ligand was greatly dependent on CD8.
Mots-clé
Affinity Labels, Amino Acid Sequence, Animals, Antibodies, Monoclonal/pharmacology, Antigens, Protozoan/metabolism, Clone Cells, Edetic Acid/pharmacology, Egtazic Acid/pharmacology, Humans, Ligands, Molecular Sequence Data, Photochemistry, Plasmodium berghei, Protozoan Proteins/metabolism, Receptors, Antigen, T-Cell/antagonists &amp, inhibitors, Receptors, Antigen, T-Cell/chemistry, T-Lymphocytes, Cytotoxic/chemistry, T-Lymphocytes, Cytotoxic/immunology
Pubmed
Web of science
Création de la notice
28/01/2008 12:20
Dernière modification de la notice
20/08/2019 15:18
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