Cell-Free Assays to Measure Effects of Regulatory Ligands on AMPK

Methods in Molecular Biology
Fiona A FyffeD G Hardie

Abstract

AMP-activated protein kinase (AMPK) is an energy sensor that is activated by increases in the cellular AMP/ATP and ADP/ATP ratios by three mechanisms: (1) allosteric activation, (2) promotion of phosphorylation at Thr172 on the α subunit by upstream kinases, and (3) inhibition of dephosphorylation of Thr172 by protein phosphatases. All of these effects are triggered by the binding of AMP or ADP at one or more of three sites on the γ subunit, where they displace ATP. AMPK is also activated by ligands that bind in the ADaM site, which is located between the α and β subunits. In this chapter we describe cell-free assays that can be used to study these varied activation mechanisms.

Citations

Feb 6, 2020·Biochemical Society Transactions·Hannah CrockerImre Berger
Jan 29, 2020·Cell Chemical Biology·Simon A HawleyD Grahame Hardie
Sep 9, 2021·Proceedings of the National Academy of Sciences of the United States of America·Joshua C DrakeZhen Yan

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