Abstract
Receptor tyrosine kinase (RTK) signaling exists in equilibrium between RTK tyrosyl phosphorylation and RTK tyrosyl dephosphorylation. Despite a detailed understanding of RTK tyrosyl phosphorylation, much less is known about RTK tyrosyl dephosphorylation. The receptor PTPs (RPTPs) are outstanding targets for the dephosphorylation of RTKs because of their mutual membrane proximity. In this chapter, we describe how to identify RPTPs that modulate the activity of RTKs using a siRNA screen and commercially available proteomic applications. The validation of putative RTKs as RPTP substrates using substrate-trapping approaches is detailed.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Antibodies, Phospho-Specific / chemistry
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Cell Line
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Gene Expression Regulation
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High-Throughput Screening Assays*
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Humans
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Phosphorylation
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Protein Array Analysis / instrumentation
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Protein Tyrosine Phosphatases / genetics
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Protein Tyrosine Phosphatases / metabolism*
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism*
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Receptor Protein-Tyrosine Kinases / antagonists & inhibitors
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Receptor Protein-Tyrosine Kinases / genetics
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Receptor Protein-Tyrosine Kinases / metabolism*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism*
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Signal Transduction
Substances
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Antibodies, Phospho-Specific
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RNA, Small Interfering
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Recombinant Fusion Proteins
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Receptor Protein-Tyrosine Kinases
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Protein Tyrosine Phosphatases