Discovery of Native Protein Complexes by Liquid Chromatography Followed by Quantitative Mass Spectrometry

2021 ◽  
pp. 105-128
Author(s):  
Wasim Aftab ◽  
Axel Imhof
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Protein Complexes ◽  
Native Protein ◽  
Quantitative Mass Spectrometry

scholarly journals Ion Imaging of Native Protein Complexes Using Orthogonal Time-of-Flight Mass Spectrometry and a Timepix Detector

2021 ◽  
Vol 32 (2) ◽  
pp. 569-580
Author(s):  
Anjusha Mathew ◽  
Ronald Buijs ◽  
Gert B. Eijkel ◽  
Frans Giskes ◽  
Andrey Dyachenko ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Complexes ◽  
Time Of Flight ◽  
Ion Imaging ◽  
Native Protein ◽  
Flight Mass Spectrometry

scholarly journals Identifying specific protein interaction partners using quantitative mass spectrometry and bead proteomes

2008 ◽  
Vol 183 (2) ◽  
pp. 223-239 ◽  
Author(s):  
Laura Trinkle-Mulcahy ◽  
Séverine Boulon ◽  
Yun Wah Lam ◽  
Roby Urcia ◽  
François-Michel Boisvert ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Interaction ◽  
Cell Biology ◽  
Protein Complexes ◽  
Affinity Purification ◽  
Specific Protein ◽  
Protein Interaction Data ◽  
Quantitative Mass Spectrometry ◽  
Cell Extracts ◽  
Interaction Partners

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.

scholarly journals Stoichiometry of chromatin-associated protein complexes revealed by label-free quantitative mass spectrometry-based proteomics

2012 ◽  
Vol 41 (1) ◽  
pp. e28-e28 ◽  
Author(s):  
Arne H. Smits ◽  
Pascal W. T. C. Jansen ◽  
Ina Poser ◽  
Anthony A. Hyman ◽  
Michiel Vermeulen
Keyword(s):  
Mass Spectrometry ◽  
Protein Complexes ◽  
Label Free ◽  
Quantitative Mass Spectrometry

scholarly journals Analysis of Human Nuclear Protein Complexes by Quantitative Mass Spectrometry Profiling

PROTEOMICS ◽  
2018 ◽  
Vol 18 (11) ◽  
pp. 1700427 ◽  
Author(s):  
Katelyn E. Connelly ◽  
Victoria Hedrick ◽  
Tiago Jose Paschoal Sobreira ◽  
Emily C. Dykhuizen ◽  
Uma K. Aryal
Keyword(s):  
Mass Spectrometry ◽  
Nuclear Protein ◽  
Protein Complexes ◽  
Quantitative Mass Spectrometry
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