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Interaction of the adaptor protein Shc and the adhesion molecule cadherin
1Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
Abstract:
In mitogenic signaling pathways, Shc participates in the growth factor activation of Ras by interacting with activated receptors and/or the Grb-2.Sos complex. Using several experimental approaches we demonstrate that Shc, through its SH2 domain, forms a complex with the cytoplasmic domain of cadherin, a transmembrane protein involved in the Ca2+-dependent regulation of cell-cell adhesion. This interaction is demonstrated in a yeast two-hybrid assay, by co-precipitation from mammalian cells, and by direct biochemical analysis in vitro. The Shc-cadherin association is phosphotyrosine-dependent and is abrogated by addition of epidermal growth factor to A-431 cells maintained in Ca2+-free medium, a condition that promotes changes in cell shape. Shc may therefore participate in the control of cell-cell adhesion as well as mitogenic signaling through Ras.
Insights
The study reveals that Shc protein directly binds to cadherin, a cell adhesion molecule. This interaction, dependent on phosphotyrosine, suggests Shc influences both cell adhesion and growth factor signaling pathways.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Shc protein is known to mediate growth factor signaling by interacting with activated receptors and the Grb-2.Sos complex to activate Ras.
- Cadherin is a transmembrane protein crucial for calcium-dependent cell-cell adhesion and regulation.
Purpose of the Study:
- To investigate the potential interaction between Shc protein and cadherin.
- To elucidate the functional implications of Shc-cadherin association in cellular processes.
Main Methods:
- Yeast two-hybrid assay to detect protein-protein interactions.
- Co-precipitation experiments in mammalian cells.
- In vitro biochemical analysis to confirm direct binding.
Main Results:
- Demonstrated a direct physical complex formation between Shc and the cytoplasmic domain of cadherin.
- Confirmed the Shc-cadherin association is dependent on phosphotyrosine.
- Observed that epidermal growth factor abrogates this interaction under specific cell culture conditions.
Conclusions:
- Shc protein interacts with cadherin, suggesting a role in regulating cell-cell adhesion.
- This interaction implies Shc may integrate signals between cell adhesion and mitogenic pathways involving Ras.