Neuroscience 2005 Abstract
| Presentation Number: | 133.10 |
|---|---|
| Abstract Title: | Notch interacts with app: a bimolecular fluorescence complementation study. |
| Authors: |
Abraham, C. R.*1
; Oh, S.1
; Chen, C.1
1Dept Biochem, Boston Univ. School of Medicine, Boston, MA |
| Primary Theme and Topics |
Disorders of the Nervous System - Neurodegenerative and Movement Disorders -- Alzheimer's disease: APP and presenilin -- other mechanisms |
| Session: |
133. Presenilins and APP Proteases I Slide |
| Presentation Time: | Sunday, November 13, 2005 10:15 AM-10:30 AM |
| Location: | Washington Convention Center - Room 151B |
| Keywords: | Alzheimer's disease, Signaling, Development, Gamma secretase |
In order to decipher the physiologic function of APP, we searched for molecules that interact with the plasma membrane APP. Rat primary neurons were infected with a viral vector encoding APP751 and potentially interacting proteins were crosslinked to APP. The resulting APP complexes were immunopurified with APP antibodies conjugated to beads, the purified complexes were in-gel digested, and peptides were identified by mass spectrometry. Notch2 was identified as an APP-interacting protein (Oh et al., 2004, SFN Abstract). To confirm this surprising result, we performed co-immunoprecipitation studies in HEK293 cells stably overexpressing APP751 and transiently expressing several Notch2 or Notch1 constructs. Both full-length Notch proteins and a number of deletion mutants were able to bring down APP and a high molecular weight complex containing APP and Notch, as demonstrated by western blot. To illustrate the subcellular localization of these two transmembrane proteins, COS cells were transiently transfected with APP-CFP and Notch-YFP. As previously described, both proteins are found on the plasma membrane and in intracellular compartments. In order to prove that Notch and APP not only co-localize but also interact with each other, new APP and Notch constructs were made that contained the N-terminal or the C-terminal half of the YFP molecule, respectively. Only if APP and Notch interact, the two halves of YFP will complement each other and the complex will become fluorescent. Using bimolecular fluorescence complementation (BiFC), we were able to provide evidence that APP and Notch interact on the plasma membrane and in intracellular compartments. In addition, we show that in contrast to APP that can form homodimers, Notch proteins did not dimerize. The APP-Notch interactions may have important implications for normal development as well as for aging and neurodegenerative diseases.
Supported by AG00001 and private donations.
Sample Citation:
[Authors]. [Abstract Title]. Program No. XXX.XX. 2005 Neuroscience Meeting Planner. Washington, DC: Society for Neuroscience, 2005. Online.
Copyright © 2005-2026 Society for Neuroscience; all rights reserved. Permission to republish any abstract or part of any abstract in any form must be obtained in writing by SfN office prior to publication.
