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AMPK

Methods and Protocols

  • Book
  • © 2018

Overview

Editors:
  1. Dietbert Neumann

    1. Department of Pathology CARIM School for Cardiovascular Diseases, Maastricht University, Maastricht, The Netherlands

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  2. Benoit Viollet

    1. Department EMD, Inserm, U1016, Institut Cochin, CNRS, UMR8104, Université Paris Descartes, Paris, France

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    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 1732)

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Table of contents (37 protocols)

  1. Front Matter

    Pages i-xvii
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  2. Production and Crystallization of Full-Length Human AMP-Activated Protein Kinase (α1β1γ1)

    • Julia A. Hubbard, Bing Xiao, Jon R. Wilson
    Pages 1-14

  3. 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

  4. 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

  5. Biochemical Measurement of Glycogen: Method to Investigate the AMPK-Glycogen Relationship

    • Elite Possik, Arnim Pause
    Pages 57-67

  6. 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

  7. 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

  8. Bioinformatics Approach to Identify Novel AMPK Targets

    • Brendan Gongol, Traci Marin, David A. Johnson, John Y.-J. Shyy
    Pages 99-109

  9. Studying AMPK in an Evolutionary Context

    • Arpit Jain, Valentin Roustan, Wolfram Weckwerth, Ingo Ebersberger
    Pages 111-142

  10. AMPK Protein Interaction Analyses by Yeast Two-Hybrid

    • Pascual Sanz, Rosa Viana, Maria Adelaida Garcia-Gimeno
    Pages 143-157

  11. Transient Expression of AMPK Heterotrimer Complexes in Mammalian Cells

    • Jonathan S. Oakhill, John W. Scott, Toby A. Dite
    Pages 159-169

  12. 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

  13. Compound C/Dorsomorphin: Its Use and Misuse as an AMPK Inhibitor

    • Biplab Dasgupta, William Seibel
    Pages 195-202

  14. Identifying the Heterotrimeric Complex Stoichiometry of AMPK in Skeletal Muscle by Immunoprecipitation

    • Jesper B. Birk, Jørgen F. P. Wojtaszewski
    Pages 203-213

  15. Kinase Activity Determination of Specific AMPK Complexes/Heterotrimers in the Skeletal Muscle

    • Jesper B. Birk, Jørgen F. P. Wojtaszewski
    Pages 215-228

  16. Determination of Adenine Nucleotide Concentrations in Cells and Tissues by High-Performance Liquid Chromatography

    • Noemí García-Tardón, Bruno Guigas
    Pages 229-237

  17. 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

  18. Cellular Application of Genetically Encoded Sensors and Impeders of AMPK

    • Takafumi Miyamoto, Elmer Rho, Allen Kim, Takanari Inoue
    Pages 255-272

  19. Assessing Mitochondrial Bioenergetics by Respirometry in Cells or Isolated Organelles

    • Guillaume Vial, Bruno Guigas
    Pages 273-287

  20. 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
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Keywords

About this book

This volume explores the latest methods used to study AMPK by computational, biochemical, biophysical, cellular, and ex vivo and in vivo approaches. The chapters in this book cover specific topics, such as methods to measure change in cellular energy metabolism and analyze metabolic pathways regulated by AMPK; bioinformatics tools to identify AMPK targets; knockdown of AMPK by CRISPR-Cas9; production and crystallization of full-length human AMP-activated protein kinase; cell-free assays to measure the effects of regulatory ligands on AMPK; use of sensors of AMPK activity; AMPK protein interaction by yeast two-hybrid; the role of AMPK in inflammation and autophagy; analyzing the AMPK function in C. elegans and mammals (with special focus on skeletal muscle, blood vessels, kidneys, pancreatic islets and hypothalamus); and human γ2 AMPK mutations. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting edge and thorough, AMPK: Methods and Protocols is a valuable resource for graduate students, postdoctoral researchers, and established investigators who are interested in the richly diverse AMPK field.



Editors and Affiliations

  • Department of Pathology CARIM School for Cardiovascular Diseases, Maastricht University, Maastricht, The Netherlands

    Dietbert Neumann

  • Department EMD, Inserm, U1016, Institut Cochin, CNRS, UMR8104, Université Paris Descartes, Paris, France

    Benoit Viollet

Bibliographic Information

  • Book Title: AMPK

  • Book Subtitle: Methods and Protocols

  • Editors: Dietbert Neumann, Benoit Viollet

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-7598-3

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC 2018

  • Hardcover ISBN: 978-1-4939-7597-6Published: 26 February 2018

  • Softcover ISBN: 978-1-4939-9249-2Published: 10 December 2019

  • eBook ISBN: 978-1-4939-7598-3Published: 26 February 2018

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XVII, 626

  • Number of Illustrations: 42 b/w illustrations, 72 illustrations in colour

  • Topics: Protein Science

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