Mass Spectrometry-Based Analysis of TRPP2 Phosphorylation

Methods Mol Biol. 2019;1987:51-64. doi: 10.1007/978-1-4939-9446-5_4.

Abstract

Differential phosphorylation of proteins is a key regulatory mechanism in biology. Immunoprecipitation-coupled mass spectrometry facilitates the targeted analysis of transient receptor ion potential channel polycystin-2 (TRPP2) phosphorylation. However, empirical testing is required to optimize experimental conditions for immunoprecipitation and mass spectrometry. Here, we present a detailed workflow for the reliable analysis of endogenous TRPP2 phosphorylation in differentiated renal epithelial cells.

Keywords: ADPKD; Immunoprecipitation; Mass spectrometry; PKD2; Phosphoproteomics; Phosphorylation; Polycystic kidney disease; Post-translational modification; TRPP2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Epithelial Cells / enzymology
  • Kidney / enzymology
  • Mass Spectrometry / methods*
  • Mice
  • Phosphorylation
  • TRPP Cation Channels / chemistry
  • TRPP Cation Channels / metabolism*

Substances

  • TRPP Cation Channels
  • polycystic kidney disease 2 protein