Recombinant cAMP-dependent protein kinase: PKA, holo type II alpha, 25 µg

Background: cAMP-dependent protein kinase (PKA) is an ubiquitous serine/theonine protein kinase present in a variety of tissues (e.g. brain, skeletal muscle, heart). The intracellular cAMP level regulates cellular responses by altering the interaction between the catatytic C and regulatory R subunits of PKA. The inactive tetrameric PKA holoenzyme R2C2 is activated when cAMP binds to R2, which dissociates the tetramer to R2*cAMP4 and two active catalytic subunits. Free catalytic subunits of PKA can phosphorylate a wide variety of intracellular target proteins. In response to hormone-induced high cAMP levels, PKA phosphorylates glycogen synthetase (inhibition of the enzyme activity) and phosphorylase kinase to block glycogen synthesis.

Inactive holoenzyme consisting of one dimeric regulatory subunit type IIα (monomer 45 kD) and two monomeric catalytic subunits(cAMP-free, monomer of 41 kD). Holo enzymecan be activated by adding the second messenger cAMP (activation constant about 100 nM) releasing two monomeric catalytic subunits. The product is suitable for analysing PKA type II agonists (cAMP analogs) or antagonists.

Theoretical MW: 172 kDA
Expression system: E. coli
Storage buffer: 20 mM MOPS (pH 7.0), 150 mM NaCl, 50% glycerol
Purity: >95% (SDS-PAGE)
Protein concentration: 1.2 mg/ml (Bradford method using BSA as standard protein)

Specific Activity: The specific activity of PKA catalytic subunit upon activation of the inactive PKA holoenzyme, type IIα, is  >15,000,000 U/mg whereby one unit is defined as the amount of catalytic subunit required to incorporate 1 pmol of phosphate into the substrate peptide Kemptide (LRRASLG) in one minute at 30°C.

Ordering information: shipped on dry ice

Entrez Gene ID: 18747/5576
UniProtKB: P05132/P13861

Product Citations:

Wolter S, Golombek M, Seifert R. (2011) "Differential activation of cAMP- and cGMP-dependent protein kinases by cyclic purine and pyrimidine nucleotides."Biochem Biophys Res Commun. 2011 Dec 2;415(4):563-6.

Courilleau D, Bisserier M, Jullian JC, Lucas A, Bouyssou P, Fischmeister R, Blondeau JP, Lezoualc'h F. (2012) "Identification of a tetrahydroquinoline analog as a pharmacological inhibitor of the cAMP-binding protein Epac." J Biol Chem. 2012 Dec 28;287(53):44192-202.