Product Pathways - Glucose Metabolism
AMPKα1 (1-550) Kinase #7464
Cell Signaling Technology offers a full line of protein kinases, substrates, antibody detection reagents and HTScan® kits. Browse our "Reagents for High-Throughput Screening" product listing or contact us at drugdiscovery@cellsignal.com.
Description
Purified recombinant kinase, supplied as a GST fusion protein.
Source / Purification
This GST-kinase fusion protein was produced using a baculovirus expression system with a construct expressing the kinase with an amino terminal GST tag. The protein was purified by one-step affinity chromatography using glutathione-agarose.
Quality Control
The purified kinase fusion protein was quality controlled for purity using SDS-PAGE followed by Coomassie or silver stain and Western blot. The specific activity of the kinase was determined using a radiometric assay.
Background
AMP-activated protein kinase (AMPK) is highly conserved from yeast to plants and animals and plays a key role in the regulation of energy homeostasis (1). AMPK is a heterotrimeric complex composed of a catalytic α subunit and regulatory β and γ subunits, each of which is encoded by two or three distinct genes (α1, 2; β1, 2; γ1, 2, 3) (2). The kinase is activated by an elevated AMP/ATP ratio due to cellular and environmental stress, such as heat shock, hypoxia and ischemia (1). The tumor suppressor LKB1, in association with accessory proteins STRAD and MO25, phosphorylates AMPKα at Thr172 in the activation loop and this phosphorylation is required for AMPK activation (3-5). AMPKα is also phosphorylated at Thr258 and Ser485 (for α1; Ser491 for α2). The upstream kinase and the biological significance of these phosphorylation events have yet to be elucidated (6). The β1 subunit is post-translationally modified by myristoylation and multi-site phosphorylation including Ser24/25, Ser96, Ser101, Ser108 and Ser182 (6,7). Phosphorylation at Ser108 of the β1 subunit seems to be required for the activation of AMPK enzyme, while phosphorylation at Ser24/25 and Ser182 affects AMPK localization (7). Several mutations in AMPKγ subunits have been identified, most of which are located in the putative AMP/ATP binding sites (CBS or Bateman domains). Mutations at these sites lead to reduction of AMPK activity and cause glycogen accumulation in heart or skeletal muscle (1,2). Accumulating evidence indicates that AMPK not only regulates the metabolism of fatty acids and glycogen, but also modulates protein synthesis and cell growth through EF2 and TSC2/mTOR pathways, as well as blood flow via eNOS/nNOS (1).
- Hardie, D.G. (2004) J. Cell Sci. 117, 5479-5487.
- Carling, D. (2004) Trends Biochem. Sci. 29, 18-24.
- Hawley, S.A. et al. (1996) J. Biol. Chem. 271, 27879-27887.
- Lizcano, J.M. et al. (2004) EMBO J. 23, 833-843.
- Shaw, R. et al. (2004) Proc. Natl. Acad. Sci. USA 101, 3329-3335.
- Woods, A. et al. (2003) J. Biol. Chem. 278, 28434-28442.
- Warden, S.M. et al. (2001) Biochem. J. 354, 275-283.
Application References
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Companion Products
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
