Transient expression of AMPK heterotrimer complexes in mammalian cells

Jonathan S. Oakhill, John W. Scott, Toby A. Dite

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

2 Citations (Scopus)


Regulation of AMP-activated protein kinase (AMPK) signalling is complex and involves contributions from adenine nucleotides, co-/posttranslational modifications, and isoform composition of the AMPK heterotrimer. It is becoming apparent that AMPK activation/inhibition by synthetic drugs involves similar levels of complexity. Major advances in our understanding of these mechanisms have been gained from recombinant expression systems that provide sufficient quantities of highly purified material for structure/function studies. Here, we provide a detailed protocol for transient expression of affinity-tagged AMPK complexes in mammalian cells. We have found this system to be optimal as a source of enzyme possessing regulatory modifications found in vivo.

Original languageEnglish
Title of host publicationAMPK
Subtitle of host publicationMethods and Protocols
EditorsDietbert Neumann, Benoit Viollet
PublisherHumana Press
Number of pages11
ISBN (Electronic)9781493975983
ISBN (Print)9781493975976
Publication statusPublished - 2018
Externally publishedYes

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • Biochemistry
  • Kinase
  • Mammalian cell
  • Metabolism
  • Purification
  • Recombinant protein
  • Transfection

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