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Visualization of APP dimerization and APP-Notch2 heterodimerization in living cells using bimolecular fluorescence
Ci-Di Chen1, Sun-Young Oh, Jason D Hinman
1Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
Journal of Neurochemistry
|March 7, 2006
Summary
Amyloid precursor protein (APP) interacts with Notch1 in rat brains. Using bimolecular fluorescence complementation, researchers visualized APP and Notch2 interactions, revealing heterodimers at the cell membrane and ER.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- The amyloid precursor protein (APP) is implicated in various cellular processes.
- Notch receptors play critical roles in cell-cell communication and development.
- Previous studies suggested an interaction between APP and Notch receptors.
Purpose of the Study:
- To confirm the endogenous interaction between APP and Notch1 in vivo.
- To investigate homodimeric and heterodimeric interactions of APP and Notch2 (N2).
- To visualize the subcellular localization of APP/N2 complexes in living cells.
Main Methods:
- Endogenous APP/Notch1 interaction confirmed in embryonic day 17 rat brain tissue.
- Bimolecular fluorescence complementation (BiFC) assay used to visualize APP/N2 interactions in COS-7 cells.
- Yellow fluorescent protein (YFP) fragments fused to APP, full-length N2, and truncated N2.
Main Results:
- APP/Notch1 endogenous interaction validated, ruling out overexpression artifacts.
- APP homodimers showed weak fluorescence; N2 did not form homodimers.
- Strongest fluorescence observed for APP/truncated N2 heterodimers, localized to plasma membrane, ER, and Golgi.
Conclusions:
- APP and Notch receptors interact endogenously.
- APP and Notch2 form heterodimers, not homodimers.
- BiFC is a valuable tool for studying APP/Notch signaling in development and disease.