Affinity methods for phosphorylation-dependent interactions.

Greg Moorhead, Carol MacKintosh

Research output: Contribution to journalReview articlepeer-review

4 Citations (Scopus)

Abstract

14-3-3s are a highly conserved protein family that exert many regulatory roles in eukaryotic cells by binding to phosphopeptide motifs in diverse target proteins. Here, we describe 14-3-3 affinity binding procedures that can be used to purify and identify 14-3-3-binding phosphoproteins; monitor how their phosphorylation and 14-3-3 binding is regulated by extracellular stimuli; define the functional effects of 14-3-3s on individual targets; and identify relevant protein phosphatases and kinases. In principle, these methods could be adapted to characterize other types of protein-protein interaction that depend on covalent modification of target sites.

Original languageEnglish
Pages (from-to)469-478
Number of pages10
JournalMethods in Molecular Biology (Clifton, N.J.)
Volume261
DOIs
Publication statusPublished - 1 Jan 2004

Keywords

  • Protein-protein interactions
  • affinity chromatography
  • phosphorylation
  • phosphopeptide

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