S.Y. Oh and A. Ellenstein contributed equally to this article.
Amyloid precursor protein interacts with notch receptors
Article first published online: 23 AUG 2005
Copyright © 2005 Wiley-Liss, Inc.
Journal of Neuroscience Research
Volume 82, Issue 1, pages 32–42, 1 October 2005
How to Cite
Oh, S. Y., Ellenstein, A., Chen, C.-D., Hinman, J. D., Berg, E. A., Costello, C. E., Yamin, R., Neve, R. L. and Abraham, C. R. (2005), Amyloid precursor protein interacts with notch receptors. J. Neurosci. Res., 82: 32–42. doi: 10.1002/jnr.20625
- Issue published online: 21 SEP 2005
- Article first published online: 23 AUG 2005
- Manuscript Accepted: 11 JUL 2005
- Manuscript Revised: 27 JUN 2005
- Manuscript Received: 1 MAY 2005
- NIH. Grant Numbers: AG00001, P41 RR10888, S10 RR15942
- Alzheimer's disease;
- transmembrane receptors;
- APP processing
The amyloid precursor protein (APP) must fulfill important roles based on its sequence conservation from fly to human. Although multiple functions for APP have been proposed, the best-known role for this protein is as the precursor of Aβ peptide, a neurotoxic 39–43-amino acid peptide crucial to the pathogenesis of Alzheimer's disease. To investigate additional roles for APP with an eye toward understanding the molecular basis of the pleiotropic effects ascribed to APP, we isolated proteins that interacted with the plasma membrane isoform of APP. We employed a membrane-impermeable crosslinker to immobilize proteins binding to transmembrane APP in human embryonic kidney (HEK)293 cells expressing APP751 (HEK275) or rat embryonic day 18 primary neurons infected with a virus expressing APP. Notch2 was identified as a potential APP binding partner based on mass spectrometry analysis of APP complexes immunopurified from neurons. To confirm the interaction between Notch2 and APP, we carried out immunoprecipitation studies in HEK275 cells transiently expressing full-length Notch2 using Notch2 antibodies. The results indicated that APP and Notch2 interact in mammalian cells, and confirmed our initial findings. Interestingly, Notch1 also coimmunoprecipitated with APP, suggesting that APP and Notch family members may engage in intermolecular cross talk to modulate cell function. Finally, cotransfection of APP/CFP and Notch2/YFP into COS cells revealed that these two proteins colocalize on the plasma membrane. Intracellularly, however, although some APP and Notch molecules colocalize, others reside in distinct locations. The discovery of proteins that interact with APP may aid in the identification of new functions for APP. © 2005 Wiley-Liss, Inc.