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

Abstract

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.
Pages255-272
Number of pages18
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1732
ISSN (Print)1064-3745

Keywords

  • 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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