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- W2521178765 abstract "Protein complexes are essential in all organizational and functional aspects of the cell. Different strategies currently exist for analyzing such protein complexes by mass spectrometry, including affinity purification (AP-MS) and proximal labeling-based strategies. However, the high sensitivity of current mass spectrometers typically results in extensive protein lists mainly consisting of nonspecifically copurified proteins. Finding the true positive interactors in these lists remains highly challenging. Here, we report a powerful design based on differential labeling with stable isotopes combined with nonequal mixing of control and experimental samples to discover bona fide interaction partners in AP-MS experiments. We apply this intelligent mixing of proteomes (iMixPro) concept to overexpression experiments for RAF1, RNF41, and TANK and also to engineered cell lines expressing epitope-tagged endogenous PTPN14, JIP3, and IQGAP1. For all baits, we confirmed known interactions and found a number of novel i..." @default.
- W2521178765 created "2016-09-30" @default.
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- W2521178765 date "2016-09-28" @default.
- W2521178765 modified "2023-10-17" @default.
- W2521178765 title "Intelligent Mixing of Proteomes for Elimination of False Positives in Affinity Purification-Mass Spectrometry" @default.
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- W2521178765 doi "https://doi.org/10.1021/acs.jproteome.6b00517" @default.
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