Anti-peptide antibodies detect conformational changes of the inter-SH2 domain of ZAP-70 due to binding to the zeta chain and to intramolecular interactions.

The Journal of Biological Chemistry
L GrazioliO Acuto

Abstract

T cell receptor (TCR) triggering induces association of the protein tyrosine kinase ZAP-70, via its two src-homology 2 (SH2) domains, to di-phosphorylated Immunoreceptor Tyrosine-based Activation Motifs (2pY-ITAMs) present in the intracellular tail of the TCR-zeta chain. The crystal structure of the SH2 domains complexed with a 2pY-ITAM peptide suggests that the 60-amino acid-long inter-SH2 spacer helps the SH2 domains to interact with each other to create the binding site for the 2pY-ITAM. To investigate whether the inter-SH2 spacer has additional roles in the whole ZAP-70, we raised antibodies against two peptides of this region and probed ZAP-70 structure under various conditions. We show that the reactivity of antibodies directed at both sequences was dramatically augmented toward the tandem SH2 domains alone compared with that of the entire ZAP-70. This indicates that the conformation of the inter-SH2 spacer is not maintained autonomously but is controlled by sequences C-terminal to the SH2 domains, namely, the linker region and/or the kinase domain. Moreover, antibody binding to the same two determinants was also inhibited when ZAP-70 or the SH2 domains bound to the zeta chain or to a 2pY-ITAM. Together, these two observa...Continue Reading

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Citations

Feb 6, 2002·European Journal of Immunology·Vincenzo Di BartoloOreste Acuto
May 24, 2011·The Journal of Biological Chemistry·Peter J Bond, José D Faraldo-Gómez
May 11, 2010·Cold Spring Harbor Perspectives in Biology·Haopeng WangArthur Weiss
Apr 7, 2000·Protein Science : a Publication of the Protein Society·R O'BrienJ E Ladbury
Nov 26, 2002·Immunological Reviews·Antonio S SechiMichael Probst-Kepper
Mar 18, 2009·Immunological Reviews·Byron B Au-YeungArthur Weiss
May 13, 1999·The Journal of Biological Chemistry·M PelosiO Acuto
Oct 9, 1999·Journal of Medicinal Chemistry·C B VuT K Sawyer

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