J Proteome Res 2007,
PMID: 17203973
Vasilescu, Julian; Zweitzig, Daniel R; Denis, Nicholas J; Smith, Jeffrey C; Ethier, Martin; Haines, Dale S; Figeys, Daniel
Mass spectrometry (MS) coupled to affinity purification is a powerful approach for identifying protein-protein interactions and for mapping post-translational modifications. Prior to MS analysis, affinity-purified proteins are typically separated by gel electrophoresis, visualized with a protein stain, excised, and subjected to in-gel digestion. An inherent limitation of this series of steps is the loss of protein sample that occurs during gel processing. Although methods employing in-solution digestion have been reported, they generally suffer from poor reaction kinetics. In the present study, we demonstrate an application of a microfluidic processing device, termed the Proteomic Reactor, for enzymatic digestion of affinity-purified proteins for liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. Use of the Proteomic Reactor enabled the identification of numerous ubiquitinated proteins in a human cell line expressing reduced amounts of the ubiquitin-dependent chaperone, valosin-containing protein (VCP). The Proteomic Reactor is a novel technology that facilitates the analysis of affinity-purified proteins and has the potential to aid future biological studies.
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Manually curated Databases
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IRef Biogrid Interaction:
SLC25A5
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UBC
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PSMC2
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FGFR4
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HSP90AB1
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PABPC1
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UBC
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TRAP1
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UBC
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UBE2T
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RPL13
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HSP90AA1
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UBC
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UBE2D3
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UBC
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PSMA1
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PCM1
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WDR61
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SLC25A4
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NUDT6
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UBA52
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SYNE1
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In total, 67 gene pairs are associated to this article in curated databases