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A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
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Biochemistry
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JoVE Journal Biochemistry
A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

DOI:
10.3791/63805-v

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10:17 min

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April 29, 2022

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1Department of Biochemistry and Cell Biology,Geisel School of Medicine at Dartmouth, 2Norris Cotton Cancer Center

Chapters

  • 00:04Introduction
  • 00:46Sample Preparation
  • 02:26Preparation of Phosphatase Inhibitor Beads
  • 04:03Washing and Elution of PPPs from the PIBs
  • 05:40Data Analysis
  • 08:30Results: Identification of Specific PIB Binders in HeLa Cells
  • 09:37Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol for the enrichment of endogenous phosphoprotein phosphatases and their interacting proteins from cells and tissues and their identification and quantification by mass spectrometry-based proteomics.

Tags

Keywords: Phosphoprotein PhosphatasesPIB-MSMass SpectrometryProteomicsPhosphatase InteractorsCell LysisProtein QuantificationPhosphatase Inhibitor ControlPhosphatase Inhibitor Beads

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