Crystal Structures of the Adenylate Sensor from Fission Yeast AMP-Activated Protein Kinase

The 5′-AMP (adenosine monophosphate)–activated protein kinase (AMPK) coordinates metabolic function with energy availability by responding to changes in intracellular ATP (adenosine triphosphate) and AMP concentrations. Here, we report crystal structures at 2.9 and 2.6 Å resolution for ATP- and AMP-bound forms of a core αβγ adenylate-binding domain from the fission yeast AMPK homolog. ATP and AMP bind competitively to a single site in the γ subunit, with their respective phosphate groups positioned near function-impairing mutants. Unexpectedly, ATP binds without counterions, amplifying its electrostatic effects on a critical regulatory region where all three subunits converge.

[1]  H. Watkins,et al.  Mutations in the gamma(2) subunit of AMP-activated protein kinase cause familial hypertrophic cardiomyopathy: evidence for the central role of energy compromise in disease pathogenesis. , 2001, Human molecular genetics.

[2]  L. Fananapazir,et al.  Identification of a gene responsible for familial Wolff-Parkinson-White syndrome. , 2001, The New England journal of medicine.

[3]  D. Hardie,et al.  Glucose repression/derepression in budding yeast: SNF1 protein kinase is activated by phosphorylation under derepressing conditions, and this correlates with a high AMP:ATP ratio , 1996, Current Biology.

[4]  V. Lumbreras,et al.  Domain fusion between SNF1‐related kinase subunits during plant evolution , 2001, EMBO reports.

[5]  W A Hendrickson,et al.  Selenomethionyl proteins produced for analysis by multiwavelength anomalous diffraction (MAD): a vehicle for direct determination of three‐dimensional structure. , 1990, The EMBO journal.

[6]  L. Tong,et al.  Crystal structure of the protein kinase domain of yeast AMP-activated protein kinase Snf1. , 2005, Biochemical and biophysical research communications.

[7]  H. Lodish,et al.  A Revised Model for AMP-activated Protein Kinase Structure , 2006, Journal of Biological Chemistry.

[8]  Y. Hayashizaki,et al.  Solution structure of the kinase‐associated domain 1 of mouse microtubule‐associated protein/microtubule affinity‐regulating kinase 3 , 2006, Protein science : a publication of the Protein Society.

[9]  D. Hardie,et al.  AMP-activated protein kinase: ancient energy gauge provides clues to modern understanding of metabolism. , 2005, Cell metabolism.

[10]  B. Kemp,et al.  Intrasteric control of AMPK via the γ1 subunit AMP allosteric regulatory site , 2004 .

[11]  D. Hardie Printed in U.S.A. Copyright © 2003 by The Endocrine Society doi: 10.1210/en.2003-0982 Minireview: The AMP-Activated Protein Kinase Cascade: The Key Sensor of Cellular Energy Status , 2022 .

[12]  D. Hardie,et al.  AMP‐activated protein kinase – development of the energy sensor concept , 2006, The Journal of physiology.

[13]  Galina Polekhina,et al.  Structural basis for glycogen recognition by AMP-activated protein kinase. , 2005, Structure.

[14]  R. Dutzler,et al.  Nucleotide recognition by the cytoplasmic domain of the human chloride transporter ClC-5 , 2007, Nature Structural &Molecular Biology.

[15]  L. Goodyear,et al.  AMP-activated protein kinase regulation and action in skeletal muscle during exercise. , 2001, Biochemical Society transactions.

[16]  J. Wojtaszewski,et al.  Role of AMPK in skeletal muscle metabolic regulation and adaptation in relation to exercise , 2006, The Journal of physiology.

[17]  D J Campbell,et al.  AMP-activated protein kinase, super metabolic regulator. , 2001, Biochemical Society transactions.

[18]  Supplemental Text , 2008 .

[19]  D. Hardie,et al.  CBS domains form energy-sensing modules whose binding of adenosine ligands is disrupted by disease mutations. , 2004, The Journal of clinical investigation.

[20]  J. Seidman,et al.  Constitutively active AMP kinase mutations cause glycogen storage disease mimicking hypertrophic cardiomyopathy. , 2002, The Journal of clinical investigation.

[21]  B. Kemp,et al.  Post-translational modifications of the beta-1 subunit of AMP-activated protein kinase affect enzyme activity and cellular localization. , 2001 .

[22]  J. Seger,et al.  Novel PRKAG2 Mutation Responsible for the Genetic Syndrome of Ventricular Preexcitation and Conduction System Disease With Childhood Onset and Absence of Cardiac Hypertrophy , 2001, Circulation.

[23]  T. Leff AMP-activated protein kinase regulates gene expression by direct phosphorylation of nuclear proteins. , 2001, Biochemical Society transactions.

[24]  A. Joachimiak,et al.  Characteristics and crystal structure of bacterial inosine-5'-monophosphate dehydrogenase. , 1999, Biochemistry.

[25]  S. Uchida,et al.  Adiponectin stimulates glucose utilization and fatty-acid oxidation by activating AMP-activated protein kinase , 2002, Nature Medicine.

[26]  Young-Bum Kim,et al.  Leptin stimulates fatty-acid oxidation by activating AMP-activated protein kinase , 2002, Nature.

[27]  D. Carling,et al.  Functional Analysis of Mutations in the γ2 Subunit of AMP-activated Protein Kinase Associated with Cardiac Hypertrophy and Wolff-Parkinson-White Syndrome* , 2002, The Journal of Biological Chemistry.

[28]  R. Dutzler,et al.  Crystal structure of the cytoplasmic domain of the chloride channel ClC-0. , 2006, Structure.

[29]  B. Kemp,et al.  AMP-activated Protein Kinase β Subunit Tethers α and γ Subunits via Its C-terminal Sequence (186–270)* , 2005, Journal of Biological Chemistry.

[30]  M. Carlson,et al.  Subcellular localization of the Snf1 kinase is regulated by specific beta subunits and a novel glucose signaling mechanism. , 2001, Genes & development.

[31]  M. Lazar,et al.  Regulation of Fasted Blood Glucose by Resistin , 2004, Science.

[32]  S. Berger,et al.  Structure and dimerization of the kinase domain from yeast Snf1, a member of the Snf1/AMPK protein family. , 2006, Structure.

[33]  Slawomir K. Grzechnik,et al.  Crystal structure of a tandem cystathionine‐β‐synthase (CBS) domain protein (TM0935) from Thermotoga maritima at 1.87 Å resolution , 2004, Proteins.

[34]  M van Heel,et al.  Structure of the AAA ATPase p97. , 2000, Molecular cell.

[35]  J. Scott,et al.  Yeast SNF1 is functionally related to mammalian AMP-activated protein kinase and regulates acetyl-CoA carboxylase in vivo. , 1994, The Journal of biological chemistry.