Imperial College London
Alternate

ProfessorDavidCarling

Faculty of Medicine, Institute of Clinical Sciences

Professor of Biochemistry
 
 
 
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Contact

 

+44 (0)7590 250 559david.carling

 
 
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Location

 

2.14DLMS BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Willows:2017:10.1042/BCJ20170458,
author = {Willows, R and Sanders, MJ and Xiao, B and Patel, BR and Martin, SR and Wilson, JR and Hubbard, J and Gamblin, SJ and Carling, D},
doi = {10.1042/BCJ20170458},
journal = {Biochemical Journal},
pages = {3059--3073},
title = {Phosphorylation of AMPK by upstream kinases is required for activity in mammalian cells},
url = {http://dx.doi.org/10.1042/BCJ20170458},
volume = {474},
year = {2017}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - AMP-activated protein kinase (AMPK) plays a major role in regulating metabolism andhas attracted significant attention as a therapeutic target for treating metabolic disorders.AMPK activity is stimulated more than 100-fold by phosphorylation of threonine 172(Thr172). Binding of AMP to the γ subunit allosterically activates the kinase. Additionally,many small molecules, e.g. 991, have been identified that bind between the kinasedomain and the carbohydrate-binding module of the β subunit, stabilising their interactionand leading to activation. It was reported recently that non-phosphorylated Thr172 AMPKis activated by AMP and A769662. We present here the crystal structure of non-phosphorylatedThr172 AMPK in complex with AMP and 991. This structure reveals that theactivation loop, as well as the complex overall, is similar to the Thr172 phosphorylatedcomplex. We find that in the presence of AMP and 991 non-phosphorylated Thr172,AMPK is much less active than the Thr172 phosphorylated enzyme. In human cells, thebasal level of Thr172 phosphorylation is very low (∼1%), but is increased 10-fold by treatmentwith 2-deoxyglucose. In cells lacking the major Thr172 kinases, LKB1 and CaMKKβ,Thr172 phosphorylation is almost completely abolished, and AMPK activity is virtuallyundetectable. Our data show that AMP and 991 binding to non-phosphorylated Thr172AMPK can induce an ordered, active-like, conformation of the activation loop explaininghow AMPK activity can be measured in vitro without Thr172 phosphorylation. However, ina cellular context, phosphorylation of Thr172 is critical for significant activation of AMPK.
AU  - Willows,R
AU  - Sanders,MJ
AU  - Xiao,B
AU  - Patel,BR
AU  - Martin,SR
AU  - Wilson,JR
AU  - Hubbard,J
AU  - Gamblin,SJ
AU  - Carling,D
DO  - 10.1042/BCJ20170458
EP  - 3073
PY  - 2017///
SN  - 1470-8728
SP  - 3059
TI  - Phosphorylation of AMPK by upstream kinases is required for activity in mammalian cells
T2  - Biochemical Journal
UR  - http://dx.doi.org/10.1042/BCJ20170458
UR  - http://hdl.handle.net/10044/1/50091
VL  - 474
ER  -
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