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
Authors | Jon Oakhill, John Scott and Toby A. Dite |
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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. |
Keywords | recombinant protein; linase; mammalian cell; transfection; purification; biochemistry; metabolism |
Page range | 159-169 |
Year | 2018 |
Book title | AMPK |
Publisher | Humana Press, Inc. |
Place of publication | New York |
Series | Methods in Molecular Biology |
ISBN | 9781493975983 |
ISSN | 1064-3745 |
Digital Object Identifier (DOI) | https://doi.org/10.1007/978-1-4939-7598-3_10 |
Scopus EID | 2-s2.0-85042713217 |
Publication process dates | |
Deposited | 13 May 2021 |
https://acuresearchbank.acu.edu.au/item/8w0w1/transient-expression-of-ampk-heterotrimer-complexes-in-mammalian-cells
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