Figure 1. MAPKAPK-5 kinase activity was measured in a radiometric assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 3 mM MgCl2, 3 mM MnCl2, 3 µM Na-orthovanadate, 1.2 mM DTT, ATP, variable, 2.5 µg/50 µl PEG20.000, Substrate: Tetra (LRRWSLG), 3 µg/50 µl and recombinant MAPKAPK-5: 200 ng/50 µl.
The kit provides a means of performing kinase activity assays with recombinant human MAPKAPK-5 kinase. It includes active MAPKAPK-5 kinase (supplied as a GST fusion protein), a biotinylated peptide substrate and a phospho-serine/threonine antibody for detection of the phosphorylated form of the substrate peptide.
Peptide substrate, Biotin- Tyrosine Hydroxylase (Ser40): 2,326 Daltons. GST-MAPKAPK-5 Kinase: 84 kDa.
MAPKAPK-5 belongs to the mitogen-activated protein kinase (MAPK) activated protein kinases (MK) subfamily that includes MAPKAPK-2/MK2 and MK3/3pK. The MK subfamily is part of a family of protein kinase subfamilies downstream of MAPK pathways and includes ribosomal S6 kinase (RSKs), mitogen and stress activated kinases (MSKs) and MAPK-interacting kinases (MNKs). All MKs are activated by MAPK pathways and mediate important processes (e.g. gene expression, cell cycle progression) and have been implicated in inflammation and cancer (1,2). MAPKAPK-5 shows binding to and activation by p38 MAPK and extracellular-regulated kinases (Erk) (3,4). MAPKAPK-5 was shown to be activated by Erk3 and act as a chaperone to Erk3 (5,6). While overexpressed MAPKAPK-5 shares similar substrates with MAPKAPK-2, such as HSP27 and glycogen synthase, recent work with MAPKAPK-5 knock-out mice indicates distinct substrates and functional properties (7).
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