Journal
DEVELOPMENTAL DYNAMICS
Volume 232, Issue 3, Pages 827-834Publisher
WILEY-LISS
DOI: 10.1002/dvdy.20272
Keywords
Notch; Drosophila; mass spectrometry; protein complexes
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Funding
- NCI NIH HHS [CA098402] Funding Source: Medline
- NIGMS NIH HHS [GM62931] Funding Source: Medline
- NINDS NIH HHS [NS26084] Funding Source: Medline
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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. (C) 2005 Wiley-Liss, Inc.
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