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Mouse jagged1 physically interacts with notch2 and other notch receptors. Assessment by quantitative methods
1Department of Hematology and Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, 113-8655 Japan.
The Journal of Biological Chemistry
|November 7, 1999
Summary
Mouse Jagged1 (mJagged1) physically binds to Notch receptors, including Notch2, Notch1, and Notch3. The Delta/Serrate/LAG-2 (DSL) domain is essential for this interaction, while EGF-like repeats modulate binding affinity.
Area of Science:
- Cell signaling
- Molecular biology
- Protein-protein interactions
Background:
- Delta/Serrate/LAG-2 (DSL) proteins act as ligands for Notch receptors.
- The precise physical interactions between DSL proteins and Notch receptors remain unclear.
Purpose of the Study:
- To clone mouse Jagged1 (mJagged1) and investigate its binding characteristics with Notch receptors.
- To elucidate the specific domains of mJagged1 involved in receptor interaction.
Main Methods:
- Established cell-based and solid-phase binding assays using mJagged1 fusion proteins.
- Performed Scatchard analysis and deletion mutant studies.
- Utilized Ca(2+)-dependent binding assays.
Main Results:
- mJagged1 binds to mouse Notch2 (mNotch2) on cell surfaces and in soluble forms in a Ca(2+)-dependent manner.
- Dissociation constants (Kd) were determined to be 0.4 nM (cell-based) and 0.7 nM (soluble).
- The DSL domain is the minimal binding unit, and EGF-like repeats modulate binding affinity.
Conclusions:
- mJagged1 physically interacts with mNotch2, demonstrating a high affinity.
- The DSL domain is critical for mJagged1-Notch2 binding.
- mJagged1 serves as a ligand for Notch1, Notch2, and Notch3 receptors.