Proteomic Analysis of the Notch Interactome
Recent large-scale studies have provided a global description of the interactome—the whole network of protein interactions in a cell or an organism—for several model organisms. Defining protein interactions on a proteome-wide scale has led to a better understanding of the cellular functions of many proteins, especially those that have not been studied by classical molecular genetic approaches. Here we describe the resources, methods, and techniques necessary for generation of such a proteome-scale interactome in a high throughput manner. These procedures will also be applicable to low or medium throughput focused studies aimed at understanding interactions between members of specific pathways such as Notch signaling.
Key wordsNotch signaling Proteomics Protein interactions Genetic interactions Interactome Integrated network
This work was supported by NIH grants 5R01HG003616, NS26084, and CA98402 (S.A-T.), as well as a JSPS Postdoctoral Fellowship for Research Abroad (K.H.), and generation of the clone set was supported by a grant from the National Human Genome Research Institute (NHGRI P41HG3487) to our collaborator Susan Celniker.
- 26.Blaumueller CM, Qi H, Zagouras P, Artavanis-Tsakonas S (1997) Intracellular cleavage of Notch leads to a heterodimeric receptor on the plasma membrane. Cell 90 (2):281–291Google Scholar
- 27.Hu Y, Ye Y, Fortini ME (2002) Nicastrin is required for gamma-secretase cleavage of the Drosophila Notch receptor. Dev Cell 2 (1):69–78Google Scholar