Analysis of immunoreceptor tyrosine-based activation motif (ITAM) binding to ZAP-70 by surface plasmon resonance
| Autor: | Frédéric Vély, Charles J. R. Hedgecock, Bernard Malissen, Jacques A. Nunès |
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| Rok vydání: | 1997 |
| Předmět: |
Immunology
Molecular Sequence Data Receptors Antigen T-Cell chemical and pharmacologic phenomena Peptide Biosensing Techniques Biology SH2 domain Crystallography X-Ray Lymphocyte Activation Binding Competitive src Homology Domains Immunoreceptor tyrosine-based activation motif Immunology and Allergy Humans Receptors Amino Acid Amino Acid Sequence Tyrosine Receptors Immunologic Phosphotyrosine chemistry.chemical_classification ZAP-70 Protein-Tyrosine Kinase T-cell receptor hemic and immune systems Protein-Tyrosine Kinases Amino acid chemistry Biochemistry Phosphorylation Peptides Tyrosine kinase Protein Binding |
| Zdroj: | European journal of immunology. 27(11) |
| ISSN: | 0014-2980 |
| Popis: | The signaling function of the T cell antigen receptor (TCR) is mediated via CD3 polypeptides, the cytoplasmic sequences of which bear conserved immunoreceptor tyrosine-based activation motifs (ITAM). ITAM are defined by two YxxL/I sequences separated by a six-eight amino acid long spacer. Upon antigen recognition, ITAM become phosphorylated on both tyrosine residues, creating a high affinity binding site for the tandem SH2 domains found in the protein tyrosine kinase ZAP-70. Using surface plasmon resonance, we further dissected the sequences required for the binding of ZAP-70 to each TCR-associated ITAM. First, we generated protein tyrosine phosphatase-resistant ITAM peptide analogs, in which difluorophosphonomethyl phenylalanyl (F2p) replaced both phosphotyrosines, and showed that those protein tyrosine phosphatase-resistant analogs bind ZAP-70 with high affinity, establishing a rational strategy for the design of novel pharmacological tools capable of interfering with TCR signaling function. Second, we substituted the five amino acids separating the two YxxL/I sequences of the CD3 zeta 1 ITAM with a non-peptidic linker made up of gamma-amino butyric acid units and demonstrated that the length of this intervening sequence rather than its chemical composition is essential for high affinity binding of phosphorylated ITAM to the ZAP-70 SH2 domains. |
| Databáze: | OpenAIRE |
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