AMPK: Methods and Protocols

Front Matter ....Pages i-xvii
 Production and Crystallization of Full-Length Human AMP-Activated Protein Kinase (α1β1γ1)  (Julia A. Hubbard, Bing Xiao, Jon R. Wilson)....Pages 1-14
 Visualizing AMPK Drug Binding Sites Through Crystallization of Full-Length Phosphorylated α2β1γ1 Heterotrimer  (Christopher G. Langendorf, Jonathan S. Oakhill, Bruce E. Kemp)....Pages 15-27
 Biophysical Interactions of Direct AMPK Activators  (Ravi G. Kurumbail, Graham M. West, Venkatasubramanian Dharmarajan, Kris A. Borzilleri, Jane M. Withka, Jessica Ward et al.)....Pages 29-55
 Biochemical Measurement of Glycogen: Method to Investigate the AMPK-Glycogen Relationship  (Elite Possik, Arnim Pause)....Pages 57-67
 Cell-Free Assays to Measure Effects of Regulatory Ligands on AMPK  (Fiona A. Fyffe, Simon A. Hawley, Alexander Gray, D. Grahame Hardie)....Pages 69-86
 Applications of NMR and ITC for the Study of the Kinetics of Carbohydrate Binding by AMPK β-Subunit Carbohydrate-Binding Modules  (Paul R. Gooley, Ann Koay, Jesse I. Mobbs)....Pages 87-98
 Bioinformatics Approach to Identify Novel AMPK Targets  (Brendan Gongol, Traci Marin, David A. Johnson, John Y.-J. Shyy)....Pages 99-109
 Studying AMPK in an Evolutionary Context  (Arpit Jain, Valentin Roustan, Wolfram Weckwerth, Ingo Ebersberger)....Pages 111-142
 AMPK Protein Interaction Analyses by Yeast Two-Hybrid  (Pascual Sanz, Rosa Viana, Maria Adelaida Garcia-Gimeno)....Pages 143-157
 Transient Expression of AMPK Heterotrimer Complexes in Mammalian Cells  (Jonathan S. Oakhill, John W. Scott, Toby A. Dite)....Pages 159-169
 Knockdown of Human AMPK Using the CRISPR/Cas9 Genome-Editing System  (Adrien Grenier, Pierre Sujobert, Séverine Olivier, Hélène Guermouche, Johanna Mondésir, Olivier Kosmider et al.)....Pages 171-194
 Compound C/Dorsomorphin: Its Use and Misuse as an AMPK Inhibitor  (Biplab Dasgupta, William Seibel)....Pages 195-202
 Identifying the Heterotrimeric Complex Stoichiometry of AMPK in Skeletal Muscle by Immunoprecipitation  (Jesper B. Birk, Jørgen F. P. Wojtaszewski)....Pages 203-213
 Kinase Activity Determination of Specific AMPK Complexes/Heterotrimers in the Skeletal Muscle  (Jesper B. Birk, Jørgen F. P. Wojtaszewski)....Pages 215-228
 Determination of Adenine Nucleotide Concentrations in Cells and Tissues by High-Performance Liquid Chromatography  (Noemí García-Tardón, Bruno Guigas)....Pages 229-237
 Intact Cell Assays to Monitor AMPK and Determine the Contribution of the AMP-Binding or ADaM Sites to Activation  (Simon A. Hawley, Fiona A. Fyffe, Fiona M. Russell, Graeme J. Gowans, D. Grahame Hardie)....Pages 239-253
 Cellular Application of Genetically Encoded Sensors and Impeders of AMPK  (Takafumi Miyamoto, Elmer Rho, Allen Kim, Takanari Inoue)....Pages 255-272
 Assessing Mitochondrial Bioenergetics by Respirometry in Cells or Isolated Organelles  (Guillaume Vial, Bruno Guigas)....Pages 273-287
 Study of AMPK-Regulated Metabolic Fluxes in Neurons Using the Seahorse XFe Analyzer  (Claudia Marinangeli, Jérome Kluza, Philippe Marchetti, Luc Buée, Valérie Vingtdeux)....Pages 289-305
 Investigating the Role of AMPK in Inflammation  (Sarah J. Mancini, Ian P. Salt)....Pages 307-319
 Studying the Role of AMPK in Cardiac Hypertrophy and Protein Synthesis  (Florence Mailleux, Christophe Beauloye, Jean-Luc Balligand, Sandrine Horman, Luc Bertrand)....Pages 321-342
 Assessment of AMPK-Stimulated Cellular Long-Chain Fatty Acid and Glucose Uptake  (Joost J. F. P. Luiken, Dietbert Neumann, Jan F. C. Glatz, Will A. Coumans, Dipanjan Chanda, Miranda Nabben)....Pages 343-361
 Measurement of AMPK-Induced Inhibition of Lipid Synthesis Flux in Cultured Cells  (Marc Foretz, Benoit Viollet)....Pages 363-371
 Studying the Role of AMPK in Autophagy  (Sarah Krieg, Bernhard Lüscher, Jörg Vervoorts, Marc Dohmen)....Pages 373-391
 Determining AMPK Activation via the Lysosomal v-ATPase-Ragulator-AXIN/LKB1 Axis  (Chen-Song Zhang, Mengqi Li, Yue Zong, Sheng-Cai Lin)....Pages 393-411
 Manipulation and Measurement of AMPK Activity in Pancreatic Islets  (Aida Martinez-Sanchez, Marie-Sophie Nguyen-Tu, Isabelle Leclerc, Guy A. Rutter)....Pages 413-431
 Analyzing AMPK Function in the Hypothalamus  (Patricia Seoane-Collazo, Miguel López)....Pages 433-448
 Using Ex Vivo Kidney Slices to Study AMPK Effects on Kidney Proteins  (Renee Rao, Kazuhiro Omi, Roshan Rajani, Hui Li, Nuria M. Pastor-Soler)....Pages 449-464
 A Flow Cytometry-Based Protocol to Measure Lymphocyte Viability Upon Metabolic Stress  (Sébastien Denanglaire, Tiphène Pirnay, Oberdan Leo, Fabienne Andris)....Pages 465-476
 Methods to Evaluate AMPK Regulation of Macrophage Cholesterol Homeostasis  (Nicholas D. LeBlond, Morgan D. Fullerton)....Pages 477-493
 Modulation of Vascular Function by AMPK: Assessment of NO Bioavailability and Surrogates of Oxidative Stress  (Swenja Kröller-Schön, Andreas Daiber, Eberhard Schulz)....Pages 495-506
 Measurement of Reactive Oxygen Species (ROS) and Mitochondrial ROS in AMPK Knockout Mice Blood Vessels  (Qilong Wang, Ming-Hui Zou)....Pages 507-517
 Studying the Role of AMPK in Angiogenesis  (Katrin Spengler, Silke Große, Nderim Kryeziu, Anne Knierim, Regine Heller)....Pages 519-537
 Analysis of Muscle Stem Cell Fate Through Modulation of AMPK Activity  (Marine Theret, Linda Gsaier, Sabrina Ben Larbi, Michèle Weiss-Gayet, Rémi Mounier)....Pages 539-549
 Evaluating the Role of Host AMPK in Leishmania Burden  (Diana Moreira, Jérôme Estaquier, Anabela Cordeiro-da-Silva, Ricardo Silvestre)....Pages 551-563
 Analysis of Transgenerational Phenotypes Following Acute Starvation in AMPK-Deficient C. elegans  (Emilie Demoinet, Richard Roy)....Pages 565-579
 Human γ2-AMPK Mutations  (Arash Yavari, Dhruv Sarma, Eduardo B. Sternick)....Pages 581-619
Back Matter ....Pages 621-626