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

TY - JOUR

T1 - Approaches to define antigen receptor-induced serine kinase signal transduction pathways

AU - Astoul, Emmanuelle

AU - Laurence, Arian D.

AU - Totty, Nick

AU - Beer, Sandra

AU - Alexander, Denis R.

AU - Cantrell, Doreen A.

PY - 2003/3/14

Y1 - 2003/3/14

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/0038391227

U2 - 10.1074/jbc.M211252200

DO - 10.1074/jbc.M211252200

M3 - Article

C2 - 12515807

AN - SCOPUS:0038391227

SN - 0021-9258

VL - 278

SP - 9267

EP - 9275

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 11

ER -