Transient expression of AMPK heterotrimer complexes in mammalian cells

Book chapter


Jon Oakhill, John Scott and Toby A. Dite. (2018) Transient expression of AMPK heterotrimer complexes in mammalian cells. In AMPK pp. 159-169 Humana Press, Inc.. https://doi.org/10.1007/978-1-4939-7598-3_10
AuthorsJon Oakhill, John Scott and Toby A. Dite
Abstract

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.

Keywordsrecombinant protein; linase; mammalian cell; transfection; purification; biochemistry; metabolism
Page range159-169
Year2018
Book titleAMPK
PublisherHumana Press, Inc.
Place of publicationNew York
SeriesMethods in Molecular Biology
ISBN9781493975983
ISSN1064-3745
Digital Object Identifier (DOI)https://doi.org/10.1007/978-1-4939-7598-3_10
Scopus EID2-s2.0-85042713217
Publication process dates
Deposited13 May 2021
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