介绍一个实验方法：亲和纯化+质谱技术分析果蝇中的蛋白复合物
丁香园论坛
2357
Analyzing Protein Complexes in Drosophila With Tandem Affinity Purification–Mass Spectrometry
DEVELOPMENTAL DYNAMICS 232:827–834, 2005
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.
有想看全文的可以mail我：mli_1982@sina.com
DEVELOPMENTAL DYNAMICS 232:827–834, 2005
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.
有想看全文的可以mail我：mli_1982@sina.com
