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Poster

A Genetically-Encoded FRET Sensor based on AMP-Activated Protein Kinase Reports Allosteric Kinase Activation

Abstract : AMPK is a multi-protein nanomachine that is activated by multiple, complex mechanisms, allowing fine tuning of AMPK activity in different situations of metabolic stress. Binding of adenine nucleotides to the gamma subunit plays a major role in either direct allosteric activation of AMPK or modulation of AMPK phosphorylation and dephosphorylation by upstream kinases and phosphatases. These activation mechanisms require crosstalk between AMPK subunits by a nucleotide-induced conformational switch. We have engineered an AMPK complex that allows a direct, real-time readout of the AMPK conformational state by fluorescence energy transfer (FRET). This molecular sensor confirms the exquisite sensitivity of AMPK to low micromolar concentrations of AMP, shows the exclusive ability of ATP, but not MgATP, to compete with AMP, and allows insight into the role of CBS domains for allosteric AMPK activation. It has potential applications as a tool for screening of allosteric activators of AMPK, and as a reporter of cellular energy state.
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https://hal.univ-grenoble-alpes.fr/hal-01955044
Contributeur : Sarah Hamant <>
Soumis le : vendredi 14 décembre 2018 - 10:29:57
Dernière modification le : mercredi 15 juillet 2020 - 09:10:04

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Uwe Schlattner, Martin Pelosse, Imre Berger. A Genetically-Encoded FRET Sensor based on AMP-Activated Protein Kinase Reports Allosteric Kinase Activation. 59th Annual Meeting of the Biophysical Society, Feb 2015, Blatimore, United States. Elsevier, 108 (2 suppl 1), pp.612a, 2015, Biophysical Journal. ⟨10.1016/j.bpj.2014.11.3332⟩. ⟨hal-01955044⟩

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