Product Pathways - MAPK Signaling
JNK1 Kinase #7418
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Description
Purified recombinant full length mouse JNK1 (Met1-Gln384) 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 mouse JNK1 (Met1-Gln384) (GenBank Accession No. NM_016700) 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-JNK1 fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. JNK1 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: ATF2 200 ng/μL and recombinant JNK1: variable.
Quality Control
The theoretical molecular weight of the GST-JNK1 fusion protein is 71 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. JNK1 kinase activity was determined using a radiometric assay [Fig.2].
Background
The stress-activated protein kinase/Jun-amino-terminal kinase (SAPK/JNK) is potently and preferentially activated by a variety of environmental stresses, including UV and gamma radiation, ceramides, inflammatory cytokines and in some instances, by growth factors and GPCR agonists (1-6). As with the other MAPKs, the core signaling unit is composed of a MAPKKK, typically MEKK1-4, or by one of the mixed lineage kinases (MLKs), which phosphorylate and activate MKK4-7, which then phosphorylate and activate the SAPK/JNK kinase (2). Stress signals are delivered to this cascade by small GTPases of the Rho family (Rac, Rho, cdc42) (3). Both Rac1 and cdc42 mediate the stimulation of MEKKs and MLKs (3). Alternatively, MKK4-7 can be activated by a pathway independent of small GTPases via stimulation of a member of the germinal center kinase (GCK) family (4). There are three SAPK/JNK genes with further diversification resulting from alternative splicing (3). Active SAPK/JNK dimers can translocate to the nucleus to regulate transcription through its effects on c-Jun, ATF-2 and other transcription factors (3,5).
- Davis, R.J. (1999) Biochem. Soc. Symp. 64, 1-12.
- Ichijo, H. (1999) Oncogene 18, 6087-6093.
- Kyriakis, J.M. and Avruch, J. (2001) Physiol. Rev. 81, 807-869.
- Kyriakis, J.M. (1999) J. Biol. Chem. 274, 5259-5262.
- Leppa, S. and Bohmann, D. (1999) Oncogene 18, 6158-6162.
- Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem. Sci. 23, 481-485.
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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.
