Cellular application of genetically encoded sensors and impeders of AMPK

Takafumi Miyamoto, Elmer Rho, Allen Kim, Takanari Inoue

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations


Unraveling the spatiotemporal dynamics of 5'-AMP-activated protein kinase (AMPK) signaling is necessary to bridge the gap between nutrient signaling and downstream function. Three genetically encoded Förster Resonance Energy Transfer (FRET)-based AMPK biosensors are available yielding insight into how AMPK-derived signal propagates throughout a cell in response to particular inputs. These findings, together with accumulating evidence obtained from biochemical techniques, promise to give a holistic understanding of the AMPK signaling. In this protocol, we describe the procedures and materials required for imaging intracellular AMPK activity in an organelle-specific manner, with a focus on ABKAR, a FRET-based biosensor. In addition, we introduce a novel AMPK inhibitor peptide that allows us to inhibit AMPK activity at specific subcellular compartments.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages18
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Biosensor
  • Compartmentalization
  • Förster resonance energy transfer (FRET)
  • Inhibitor peptide
  • Kinase activity monitoring
  • Organelle-specific
  • Signaling dynamics

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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