Soskic, V;
Gorlach, M;
Poznanovic, S;
Boehmer, FD;
Godovac-Zimmermann, J;
(1999)
Functional proteomics analysis of signal transduction pathways of the platelet-derived growth factor beta receptor.
Biochemistry
, 38
(6)
1757 - 1764.
10.1021/bi982093r.
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Abstract
We report efficient methods for using functional proteomics to study signal transduction pathways in mouse fibroblasts following stimulation with PDGF. After stimulation, complete cellular proteins were separated using two-dimensional electrophoresis and phosphorylated proteins were detected with anti-phosphotyrosine and anti-phosphoserine antibodies. About 260 and 300 phosphorylated proteins were detected with the anti-phosphotyrosine and anti-phosphoserine antibodies, respectively, at least 100 of which showed prominent changes in phosphorylation as a function of time after stimulation. Proteins showing major time-dependent changes in phosphorylation were subjected to in-gel digestion with trypsin and identified by mass spectroscopy using MALDI-TOF mass fingerprinting and ESI peptide sequencing. We have observed phosphorylated proteins known to be part of the PDGF signal transduction pathway such as ERK 1, serine/threonine protein kinase akt and protein tyrosine phosphatase syp, proteins such as proto-oncogene tyrosine kinase fgr previously known to participate in other signal transduction pathways, and some proteins such as plexin-like protein with no previously known function in signal transduction. Information about the phosphorylation site was obtained for proto-oncogene tyrosine kinase fgr and for cardiac alpha-actin. The methods used here have proven to be suitable for the identification of time-dependent changes in large numbers of proteins involved in signal transduction pathways.
| Type: | Article |
|---|---|
| Title: | Functional proteomics analysis of signal transduction pathways of the platelet-derived growth factor beta receptor |
| DOI: | 10.1021/bi982093r |
| Publisher version: | http://dx.doi.org/10.1021/bi982093r |
| Language: | English |
| Keywords: | protein-tyrosine-phosphatase, simian sarcoma-virus, mass-spectrometry, 2-dimensional gels, sequence databases, alpha-subunits, kinase-C, phosphorylation, identification, activation |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine |
| URI: | https://discovery.ucl.ac.uk/id/eprint/109188 |
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