Approaches to define antigen receptor-induced serine kinase signal transduction pathways

Emmanuelle Astoul, Arian D. Laurence, Nick Totty, Sandra Beer, Denis R. Alexander, Doreen A. Cantrell (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    31 Citations (Scopus)

    Abstract

    In the present report we describe the properties of a novel phospho-specific antiserum that has opened a route to the characterization of antigen receptor-activated serine kinase pathways in lymphocytes. The basis for the present work was that Ser-21 in glycogen synthase kinase 3α is robustly phosphorylated following antigen receptor triggering. We predicted accordingly that antigen receptors would also stimulate phosphorylation of other proteins with a similar sequence. To test this idea we raised an antibody against the phosphopeptide RARTSpSFAEP, where pS is a phospho-serine corresponding to the glycogen synthase kinase 3α Ser-21 sequence. The resulting antiserum was called phospho antibody for proteomics-1 (PAP-1). The present study describes the properties of PAP-1 and shows that it can reveal quite striking differences in the phospho-proteome of different cell types and is able to pinpoint new targets in important signal transduction pathways. PAP-1 was used to map protein phosphorylations regulated by the antigen receptor in T cells. One of these PAP-1-reactive proteins was purified and revealed to be a previously unrecognized target for antigen receptor signal transduction, namely an "orphan" adapter SLY (Src homology 3 (SH3) domain-containing protein expressed in lymphocytes). The use of sera detecting specific phosphorylation sites is thus proved as a powerful method for the discovery of novel downstream components of antigen receptor signals in T cells.

    Original languageEnglish
    Pages (from-to)9267-9275
    Number of pages9
    JournalJournal of Biological Chemistry
    Volume278
    Issue number11
    DOIs
    Publication statusPublished - 14 Mar 2003

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