Recombinant, human 5' AMP-activated protein kinase α1/β1/γ1 active kinase, 20 µg

5' AMP-activated protein kinase / AMPK or 5' adenosine monophosphate-activated protein kinase is a cellular energy sensor conserved in all eukaryotic cells. AMPK is a heterotrimer protein kinase consisting of a α catalytic subunit, and non-catalytic β and γ subunits. The kinase activity of AMPK is activated by the stimuli that increase the cellular AMP/ATP ratio. Phosphorylation of the catalytic α subunit by LKB1 or Ca²⁺-calmodulin-dependent protein kinase kinase β results in activation of AMPK. The kinase AMPK activates myosin by phosphorylating the non-muscle myosin regulatory light chain. It was shown that activated AMPK is in transient association with several mitotic structures. 5'-AMP-activated protein kinase catalytic subunit alpha-1 is an enzyme that in humans is encoded by the PRKAA1 gene.

The human AMPK subunits α1, β1 and γ1 were co-expressed in E.coli. The α1-subunit has a N-terminal maltose binding protein (MBP) tag. The heterotrimeric AMPK complex was purified using an amylose matrix. The catalytic subunit α1 was activated by in vitro phosphorylation using CAMKK2. CAMKK2 and impurities were removed using HiLoad 16/600 Superdex 200 prep grade for size exclusion chromatography. The MBP-tag can be removed using TEV protease.

Human AMPK-alpha1, amino acids M1-Q559, N-terminally fused to MBP-TEV cleavage site, MW: 105 838 Da
Human AMPK-beta1, amino acids M1-I270, MW: 30 382 Da
Human AMPK-gamma1, amino acids M1-P331, MW: 37 579 Da

Expression system: E.coli
Storage buffer: 200 mM NaCl; 10 % (v/v) glycerol; 1 mM PMSF; 0.5 mM TCEP; 50 mM Tris /HCl pH 8.0, Complete (Roche, 1 tablet/50 ml)
Protein concentration: 1.5 mg/ml (Bradford method using BSA as standard protein)
Specific activity: 4.300.000*/ mg (1 Unit is defined as 1 picmole phosphate transferred  to synthetic peptide AMARA (AMARAASAAALARRR) per min at 30° C)

Entrez Gene ID: 5562, 5564,  5571
UniProtKB: Q13131, Q9Y478, P54619

Ordering informations: shipped on dry ice

Product specific literature

AMP-activated protein kinase: functional, heterotrimeric complexes by co-expression of subunits in Escherichia coli. Neumann D1, Woods A, Carling D, Wallimann T, Schlattner U. (2003) "Mammalian AMP-activated protein kinase: functional, heterotrimeric complexes by co-expression of subunits in Escherichia coli. Protein Expr Purif. 2003 Aug;30(2):230-7.