Mass Spectrometry-Based Analysis of TRPP2 Phosphorylation

Verónica I Dumit; Michael Köttgen; Alexis Hofherr

doi:10.1007/978-1-4939-9446-5_4

Mass Spectrometry-Based Analysis of TRPP2 Phosphorylation

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

Authors

Verónica I Dumit¹, Michael Köttgen^{2

3}, Alexis Hofherr⁴

Affiliations

¹ Core Facility Proteomics, Center for Biological Systems Analysis (ZBSA), University of Freiburg, Freiburg, Germany.
² Renal Division, Department of Medicine, Faculty of Medicine, Medical Center, University of Freiburg, Freiburg, Germany.
³ CIBSS - Centre for Integrative Biological Signalling Studies, University of Freiburg, Freiburg, Germany.
⁴ Renal Division, Department of Medicine, Faculty of Medicine, Medical Center, University of Freiburg, Freiburg, Germany. alexis.hofherr@uniklinik-freiburg.de.

PMID: 31028673
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