Synthetic energy sensor AMPfret deciphers adenylate-dependent AMPK activation mechanism

Martin Pelosse, Cécile Cottet-Roussellea, Cécile Bidand, Aurélie Dupont, Kapil Gupta, Imre Berger, Uwe Schlattner*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

11 Citations (Scopus)
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AMP-activated protein kinase AMPK senses and regulates cellular energy state. AMPK activation by increasing AMP and ADP concentrations involves a conformational switch within the heterotrimeric complex. This is exploited here for the construction of a synthetic sensor of cellular energetics and allosteric AMPK activation, AMPfret. Based on engineered AMPK fused to fluorescent proteins, the sensor allows direct, real-time readout of the AMPK conformational state by fluorescence resonance energy transfer (FRET). AMPfret faithfully and dynamically reports the binding of AMP and ADP to AMPK γ-CBS sites, competed by Mg 2+ -free ATP. FRET signals correlate with activation of AMPK by allosteric mechanisms and protection from dephosphorylation, attributed here to specific CBS sites, but does not require activation loop phosphorylation. Moreover, AMPfret detects binding of pharmacological compounds to the AMPK α/β-ADaM site enabling activator screening. Cellular assays demonstrate that AMPfret is applicable in vivo for spatiotemporal analysis of energy state and allosteric AMPK activation.

Original languageEnglish
Article number1038
Number of pages13
JournalNature Communications
Publication statusPublished - 4 Mar 2019

Structured keywords

  • BrisSynBio
  • Bristol BioDesign Institute


  • biochemistry
  • biological techniques
  • biophysics
  • cell biology
  • drug discovery


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