Product Pathways - MAPK Signaling
MAPKAPK-3 Kinase #7490
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 full length human MAPKAPK-3 (Met1-Gln382) kinase, supplied as a GST fusion protein.
Source / Purification
The GST-Kinase fusion protein was produced using a baculovirus expression system with a construct expressing full length human MAPKAPK-3 (Met1-Gln382) (GenBank Accession No. NM_004635) with an amino-terminal GST tag. The protein was purified by one-step affinity chromatography using glutathione-agarose.
Gel Staining
Figure 1. The purity of the GST-MAPKAPK-3 fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. MAPKAPK-3 kinase activity was measured in a radiometric assay using the following reaction conditions: 4 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 0.4 mM EDTA, 4 mM MgCl2, 0.05 mM DTT, 40 ng/μl BSA, 50 μM ATP, Substrate: MBP 800 ng/μL and recombinant MAPKAPK-3: variable.
Quality Control
The theoretical molecular weight of the GST-MAPKAPK-3 fusion protein is 69 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. MAPKAPK-3 kinase activity was determined using a radiometric assay [Fig.2].
Background
MAPKAPK-3 has a single potential SH3-binding site in the proline-rich amino terminus, a putative ATP-binding site, 2 MAP kinase phosphorylation site motifs and a putative nuclear localization signal. It shares 72% nucleotide and 75% amino acid identity with MAPKAPK-2 (1). MAPKAPK-3 has been shown to be activated by growth inducers and stress stimulation of cells. In vitro studies demonstrated that Erk, p38 MAP kinase and Jun amino-terminal kinase were all able to phosphorylate and activate MAPKAPK-3, which suggested a role for this kinase as an integrative element of signaling in both mitogen and stress responses (2). This kinase was reported to interact with, phosphorylate and repress the activity of E47, which is a basic helix-loop-helix transcription factor known to be involved in the regulation of tissue-specific gene expression and cell differentiation (3). MAPKAPK-3 may also support luteal maturation through the phosphorylation and activation of the nuclear transcription factor CREB (4).
- Sithanandam, G. et al. (1996) Mol. Cell Biol. 16, 868-876.
- Ludwig, S. et al. (1996) Mol. Cell Biol. 16, 6687-6697.
- Neufeld, B. et al. (2000) J. Biol. Chem. 275, 20239-20242.
- Maizels, E.T. et al. (2001) Mol. Endocrinol. 15, 716-733.
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.
