Subcellular dynamics of protein kinase A activity visualized by FRET-based reporters

Michael D. Allen, Jin Zhang

Research output: Contribution to journalArticle

Abstract

cAMP-dependent protein kinase (PKA) mediates key cellular processes via compartmentalized activity, and the ability to track its activity in living cells should help increase our understanding of this precise regulation. Here, through systematic testing of new fluorescent proteins, we developed a new FRET-based A-kinase activity reporter (AKAR), AKAR3, with a dynamic range of 31-41%, twice that of predecessors. Visualization of PKA activity at plasma membrane, cytoplasm, nucleus, and mitochondria was achieved. Targeting AKAR3 to outer mitochondrial membrane revealed that basal PKA activity at mitochondria differs from that in the cytoplasm, indicating differential regulation of PKA activity at different subcellular locations.

Original languageEnglish (US)
Pages (from-to)716-721
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume348
Issue number2
DOIs
StatePublished - Sep 22 2006

Keywords

  • Circular permutation
  • Compartmentalization
  • FRET
  • Kinase
  • Live-cell imaging
  • Mitochondria
  • PKA

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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