Department of Biochemistry & Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada.
Nat Methods. 2012 Sep;9(9):907-9. doi: 10.1038/nmeth.2131. Epub 2012 Aug 5.
Interactomes are often measured using affinity purification-mass spectrometry (AP-MS) or yeast two-hybrid approaches, but these methods do not provide stoichiometric or temporal information. We combine quantitative proteomics and size-exclusion chromatography to map 291 coeluting complexes. This method allows mapping of an interactome to the same depth and accuracy as AP-MS with less work and without overexpression or tagging. The use of triplex labeling enables monitoring of interactome rearrangements.
相互作用组通常使用亲和纯化-质谱(AP-MS)或酵母双杂交方法进行测量,但这些方法不能提供化学计量或时间信息。我们结合定量蛋白质组学和排阻色谱法来绘制 291 个共洗脱复合物。这种方法可以在不进行过表达或标记的情况下,以更少的工作量和相同的深度和准确性将相互作用组映射到 AP-MS。三联体标记的使用可以监测相互作用组的重排。