Immunoaffinity profiling of tyrosine phosphorylation in cancer cells.
Nat Biotechnol
; 23(1): 94-101, 2005 Jan.
Article
em En
| MEDLINE
| ID: mdl-15592455
ABSTRACT
Tyrosine kinases play a prominent role in human cancer, yet the oncogenic signaling pathways driving cell proliferation and survival have been difficult to identify, in part because of the complexity of the pathways and in part because of low cellular levels of tyrosine phosphorylation. In general, global phosphoproteomic approaches reveal small numbers of peptides containing phosphotyrosine. We have developed a strategy that emphasizes the phosphotyrosine component of the phosphoproteome and identifies large numbers of tyrosine phosphorylation sites. Peptides containing phosphotyrosine are isolated directly from protease-digested cellular protein extracts with a phosphotyrosine-specific antibody and are identified by tandem mass spectrometry. Applying this approach to several cell systems, including cancer cell lines, shows it can be used to identify activated protein kinases and their phosphorylated substrates without prior knowledge of the signaling networks that are activated, a first step in profiling normal and oncogenic signaling networks.
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Base de dados:
MEDLINE
Assunto principal:
Tirosina
/
Regulação Neoplásica da Expressão Gênica
/
Proteômica
/
Neoplasias
Idioma:
En
Ano de publicação:
2005
Tipo de documento:
Article