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Direct interaction between Shc and the platelet-derived growth factor beta-receptor
1Ludwig Institute for Cancer Research, Biomedical Center, Uppsala, Sweden.
The Journal of Biological Chemistry
|May 27, 1994
Summary
Platelet-derived growth factor (PDGF) beta-receptor directly binds Shc proteins via its SH2 domain. Multiple autophosphorylation sites on the PDGF beta-receptor mediate this specific Shc binding, influencing mitogenic signaling.
Area of Science:
- Cellular signaling
- Molecular biology
- Signal transduction
Background:
- Shc proteins (p52shc, p46shc) are phosphorylated by tyrosine kinases and involved in mitogenic signaling.
- Platelet-derived growth factor (PDGF) beta-receptor activation involves autophosphorylation and interaction with SH2 domain-containing molecules.
Purpose of the Study:
- To characterize the interaction between the PDGF beta-receptor and Shc proteins.
- To identify the specific binding sites and mechanisms involved in this interaction.
Main Methods:
- Co-precipitation assays to detect protein interactions.
- Bacterial fusion protein expression to test SH2 domain binding.
- Analysis of purified receptor domains.
- Site-directed mutagenesis to assess binding site importance.
Main Results:
- PDGF beta-receptor stimulation led to Shc protein coprecipitation.
- The Shc SH2 domain directly bound autophosphorylated PDGF beta-receptor.
- Multiple PDGF beta-receptor autophosphorylation sites (Tyr-579, Tyr-740, Tyr-751, Tyr-771) mediated Shc binding.
- Mutating Tyr-579 partially impaired Shc association.
Conclusions:
- Multiple autophosphorylation sites on the PDGF beta-receptor are crucial for Shc binding.
- This interaction differs from other SH2 domain protein interactions with the PDGF beta-receptor, which typically involve a single high-affinity site.