Identifying specific protein interaction partners using quantitative mass spectrometry and bead proteomes.
Identifying specific protein interaction partners using quantitative mass spectrometry and bead proteomes.
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使用定量质谱和珠蛋白质组识别特定的蛋白质相互作用伙伴。
DOI：
10.1083/jcb.200805092
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发表时间：
2008-10-20
影响因子：
7.8
通讯作者：
Lamond, Angus
中科院分区：
文献类型：
--
作者：
Trinkle-Mulcahy, Laura;Boulon, Severine;Lam, Yun Wah;Urcia, Roby;Boisvert, Francois-Michel;Vandermoere, Franck;Morrice, Nick A.;Swift, Sam;Rothbauer, Ulrich;Leonhardt, Heinrich;Lamond, Angus
The identification of interaction partners in protein complexes is a major goal in cell biology. Here we present a reliable affinity purification strategy to identify specific interactors that combines quantitative SILAC-based mass spectrometry with characterization of common contaminants binding to affinity matrices (bead proteomes). This strategy can be applied to affinity purification of either tagged fusion protein complexes or endogenous protein complexes, illustrated here using the well-characterized SMN complex as a model. GFP is used as the tag of choice because it shows minimal nonspecific binding to mammalian cell proteins, can be quantitatively depleted from cell extracts, and allows the integration of biochemical protein interaction data with in vivo measurements using fluorescence microscopy. Proteins binding nonspecifically to the most commonly used affinity matrices were determined using quantitative mass spectrometry, revealing important differences that affect experimental design. These data provide a specificity filter to distinguish specific protein binding partners in both quantitative and nonquantitative pull-down and immunoprecipitation experiments.
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影响因子：
4.1
作者：
Al-Hakim, Abdallah K.;Zagorska, Anna;Alessi, Dario R.
通讯作者：
Alessi, Dario R.
影响因子：
4.8
作者：
Baccon, J;Pellizzoni, L;Dreyfuss, G
通讯作者：
Dreyfuss, G
影响因子：
64.5
作者：
Cox, Juergen;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
11.4
作者：
Meister, G;Fischer, U
通讯作者：
Fischer, U
影响因子：
4.8
作者：
Jones, KW;Gorzynski, K;Terns, MP
通讯作者：
Terns, MP