A fluorescent reporter of AMPK activity and cellular energy stress Academic Article uri icon

Overview

MeSH Major

  • AMP-Activated Protein Kinases

abstract

  • AMP-activated protein kinase (AMPK) is activated when the AMP/ATP ratio in cells is elevated due to energy stress. Here, we describe a biosensor, AMPKAR, that exhibits enhanced fluorescence resonance energy transfer (FRET) in response to phosphorylation by AMPK, allowing spatiotemporal monitoring of AMPK activity in single cells. We show that this reporter responds to a variety of stimuli that are known to induce energy stress and that the response is dependent on AMPK α1 and α2 and on the upstream kinase LKB1. Interestingly, we found that AMPK activation is confined to the cytosol in response to energy stress but can be observed in both the cytosol and nucleus in response to calcium elevation. Finally, using this probe with U2OS cells in a microfluidic device, we observed a very high cell-to-cell variability in the amplitude and time course of AMPK activation and recovery in response to pulses of glucose deprivation.

publication date

  • April 6, 2011

Research

keywords

  • Academic Article

Identity

Language

  • eng

PubMed Central ID

  • PMC3070961

Digital Object Identifier (DOI)

  • 10.1016/j.cmet.2011.03.006

PubMed ID

  • 21459332

Additional Document Info

start page

  • 476

end page

  • 86

volume

  • 13

number

  • 4