Characterizing PKA-mediated phosphorylation of plexin using purified proteins

Taehong Yang, Jonathan R. Terman

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


Protein phosphorylation is one of the widely used posttranslational modifications that alter protein function in vivo. We recently showed phosphorylation of Drosophila Plexin A by cyclic adenosine monophosphate (cAMP)-dependent protein kinase (PKA) and subsequent inhibition of plexin-mediated repulsive guidance. This phosphorylation occurs in the active site of the plexin GTPase-activating protein (GAP) domain, which in turn inhibits endogenous GAP activity toward Ras/Rap family small GTP-binding proteins by recruiting the phospho-serine/threonine-binding protein 14-3-3ε. Here we describe how phosphorylation of Plexin A can be detected and quantitated using an in vitro kinase assay and radioactive [γ-P32 ] adenosine 5’-triphosphate (ATP).

Original languageEnglish (US)
Pages (from-to)147-159
Number of pages13
JournalMethods in Molecular Biology
StatePublished - 2017


  • Kinase assay
  • PKA
  • Phosphorylation
  • Plexin

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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