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Structural basis for AMP binding to mammalian AMP-activated protein kinase

Abstract

AMP-activated protein kinase (AMPK) regulates cellular metabolism in response to the availability of energy and is therefore a target for type II diabetes treatment1. It senses changes in the ratio of AMP/ATP by binding both species in a competitive manner2. Thus, increases in the concentration of AMP activate AMPK resulting in the phosphorylation and differential regulation of a series of downstream targets that control anabolic and catabolic pathways1,2. We report here the crystal structure of the regulatory fragment of mammalian AMPK in complexes with AMP and ATP. The phosphate groups of AMP/ATP lie in a groove on the surface of the γ domain, which is lined with basic residues, many of which are associated with disease-causing mutations. Structural and solution studies reveal that two sites on the γ domain bind either AMP or Mg·ATP, whereas a third site contains a tightly bound AMP that does not exchange. Our binding studies indicate that under physiological conditions AMPK mainly exists in its inactive form in complex with Mg·ATP, which is much more abundant than AMP. Our modelling studies suggest how changes in the concentration of AMP ([AMP]) enhance AMPK activity levels. The structure also suggests a mechanism for propagating AMP/ATP signalling whereby a phosphorylated residue from the α and/or β subunits binds to the γ subunit in the presence of AMP but not when ATP is bound.

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Figure 1: Structure of mammalian AMPK.
Figure 2: Fluorescence titrations of mant-AMP with AMPK.
Figure 3: Front and back views of the three adenyl moieties bound to the γ domain, shown in surface representation.
Figure 4: Interactions between primarily basic residues from γ with the phosphate groups of the three bound adenyl species.

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Acknowledgements

Work in both laboratories was supported by the Medical Research Council (UK). D.C. acknowledges support from the EC. We are grateful to N. Justin and I. Taylor for MALS analysis, and S. Smerdon for discussions.

Coordinates and structure factors have been deposited in the Protein Data Bank with accession codes 2V8Q, 2V92 and 2V9J.

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Correspondence to Stephen R. Martin, David Carling or Steven J. Gamblin.

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Xiao, B., Heath, R., Saiu, P. et al. Structural basis for AMP binding to mammalian AMP-activated protein kinase. Nature 449, 496–500 (2007). https://doi.org/10.1038/nature06161

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