Measuring Gli2 Phosphorylation by Selected Reaction Monitoring Mass Spectrometry. Academic Article uri icon

Overview

abstract

  • Phosphorylation is an important mechanism by which Gli proteins are regulated. When the Hedgehog (Hh) pathway is activated, multiple serine and threonine residues of Gli2 are dephosphorylated, while at least one residue undergoes phosphorylation. These changes in phosphorylation have functional relevance for the transcriptional activity of Gli proteins, as shown by in vitro and in vivo assays on Gli mutants lacking the phosphorylated residues. Here, we describe a method of quantitatively monitoring the phosphorylation of Gli proteins by triple quadrupole mass spectrometry of Gli2 immunoprecipitated from cell lysates. This method is broadly applicable to the monitoring of phosphorylation changes of immunoprecipitated Gli proteins when the putative phosphosites are known.

publication date

  • January 1, 2015

Research

keywords

  • Kruppel-Like Transcription Factors
  • Mass Spectrometry

Identity

PubMed Central ID

  • PMC4699800

Scopus Document Identifier

  • 84946430454

Digital Object Identifier (DOI)

  • 10.1007/978-1-4939-2772-2_10

PubMed ID

  • 26179043

Additional Document Info

volume

  • 1322