Analyzing protein complexes in Drosophila with tandem affinity purification–mass spectrometry
Version of Record online: 9 FEB 2005
Copyright © 2005 Wiley-Liss, Inc.
Special Issue: Drosophila as a Model System
Volume 232, Issue 3, pages 827–834, March 2005
How to Cite
Veraksa, A., Bauer, A. and Artavanis-Tsakonas, S. (2005), Analyzing protein complexes in Drosophila with tandem affinity purification–mass spectrometry. Dev. Dyn., 232: 827–834. doi: 10.1002/dvdy.20272
- Issue online: 18 FEB 2005
- Version of Record online: 9 FEB 2005
- Manuscript Accepted: 13 OCT 2004
- Manuscript Revised: 7 OCT 2004
- Manuscript Received: 9 SEP 2004
- National Institutes of Health. Grant Numbers: NS26084, GM62931, CA098402
- mass spectrometry;
- protein complexes
We describe the application of tandem affinity purification–mass spectrometry (TAP-MS) to the study of protein complexes in Drosophila. We have constructed vectors for inducible expression of TAP-tagged fusion proteins in Drosophila cultured cells and in vivo. Using these vectors, we tagged, as a paradigm, several components of the Notch signaling pathway, isolated protein complexes containing the baits and associated proteins from cells and embryos, and identified the subunits by liquid chromatography–tandem mass spectrometry (LC-MS/MS). Several known interactions involving Notch pathway elements were confirmed, and many novel potential interactions were uncovered. For some of the novel associations, we validated the interaction genetically and biochemically. We conclude that TAP, in combination with MS, can be used as an effective method for the studies of the Drosophila proteome. Developmental Dynamics 232:827–834, 2005. © 2005 Wiley-Liss, Inc.