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cAMP-dependent protein kinase, catalytic subunit Cα
1. Description
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 R2 C2 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. Different isoforms of catalytic and regulatory subunits
suggest specific functions. The recombinant C α catalytic subunit supplied by Biaffin is a 41
kDa protein. The α -isoform is the predominant form with a broad tissue distribution and
can be used for in vitro enzymological studies of neural and hormonal signal transduction or
to phosphorylate target proteins in vivo (via microinjection) including ion channels,
transcriptional activator proteins and regulatory enzymes of glycogen metabolism.
2. Product Components
cAMP-Dependent Protein Kinase, catalytic subunit C α , is recombinantly expressed in E.coli
and purified using several classical chromatographic purification steps. The enzyme is
supplied in a buffer containing 25 mM potassium phosphate (pH 6.5), 5 mM 2mercaptoethanol, 5 mM EDTA, 150 mM NaCl, 50% glycerol. The purity is >95% (SDSPAGE).
Storage Conditions: Store the cAMP-Dependent Protein Kinase, recombinant C α catalytic
subunit, at –20 oC, where it is stable for at least six months from date of purchase. Avoid
multiple freeze-thaw cycles. Store at 4 C if thawed and if entire vial will be used within two
weeks.
Specific Activity:
The specific activity of the cAMP-Dependent Protein Kinase, recombinant C α catalytic
subunit, is >15.000.000 U/mg.
Unit Definition: One unit is defined as the amount of cAMP-Dependent Protein Kinase,
recombinant C α catalytic subunit, required to incorporate 1 pmol of phosphate into the
specific substrate peptide kemptide (LRRASLG) in one minute at 30 oC.
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