Transient expression of AMPK heterotrimer complexes in mammalian cells
Jon Oakhill John Scott Toby A. Dite
Jon Oakhill
John Scott
Toby A. Dite
Author
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
Date
2018
Type
Book chapter
Journal
Book
AMPK
Volume
Issue
Page Range
159-169
Article Number
ACU Department
Non-faculty
Mary MacKillop Institute for Health Research
Faculty of Health Sciences
Mary MacKillop Institute for Health Research
Faculty of Health Sciences