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 treatment. It senses changes in the ratio of AMP/ATP by binding both species in a competitive manner. 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 pathways. 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 gamma 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 gamma 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 alpha and/or beta subunits binds to the gamma subunit in the presence of AMP but not when ATP is bound.
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The γ-subunit includes four tandem cystathionine β synthase motifs that allow AMPK to bind adenine nucleotides and react to changes in AMP, ADP, and ATP.
“The γ-subunit allows AMPK to react to changes in the level of AMP, ADP, and ATP, as it includes four tandem cystathionine β synthase motifs that allow AMPK to bind adenine nucleotides [26].”
From AMPK, Mitochondrial Function, and Cardiovascular Disease (2020) · cites this paper as [26]
Activation loop phosphorylation increases AMPK activity by about 100-fold, while allosteric regulation changes AMPK activity up to tenfold in mammalian cells and about twofold in recombinant, bacterially produced AMPK.
“Activation loop phosphorylation increases AMPK activity by about 100-fold, while allosteric regulation changes AMPK activity up to tenfold in mammalian cells and about twofold in recombinant, bacterially produced AMPK [26,28,38,39].”
From AMPK, Mitochondrial Function, and Cardiovascular Disease (2020) · cites this paper as [26]
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- Authors
- Xiao, B., Heath, R., Saiu, P., Leiper, F. C., Leone, P., Jing, C., Walker, P. A., Haire, L., Eccleston, J. F., Davis, C. T., Martin, S. R., Carling, D., & Gamblin, S. J.
- Affiliations
- National Institute for Medical Research (GB)
- Hammersmith Hospital (GB)
- Corresponding author
- Martin, Stephen R.
- Carling, David
- ORCID
- Venue
- Nature
- Published
- 2007-09
- Volume
- 449
- Issue
- 7161
- Pages
- 496-500
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- OpenAlex
- W2033497590
- Cited by
- 591
- References
- 39
- Citations by year
- 2026: 16 · 2025: 17 · 2024: 34 · 2023: 25 · 2022: 27 · 2021: 27 · 2020: 32 · 2019: 28 · 2018: 39 · 2017: 35 · 2016: 30 · 2015: 39 · 2014: 37 · 2013: 34 · 2012: 32
- FWCI
- 12.97
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- HAL (submittedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- ENLIGHTEN (Jurnal Bimbingan dan Konseling Islam) (submittedVersion)
- Science and Technology Facilities Council (submittedVersion)
- Funders
- Medical Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolism, Diabetes, and Cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Keywords
- AMPK, AMP binding, MgATP binding, gamma subunit, AMP/ATP ratio, energy sensing, kinase activation, pathogenic variants, crystal structure, type 2 diabetes
- MeSH terms
- Adenosine Monophosphate, Animals, Binding Sites, Humans, Models, Molecular, Multienzyme Complexes, Structure-Activity Relationship, Surface Properties, Protein Serine-Threonine Kinases, Protein Structure, Tertiary, Crystallography, X-Ray, Rats, AMP-Activated Protein Kinases
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