Product Pathways - Cytoskeletal Signaling
Mst2 Kinase #7678
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 Mst2 (Met1-Phe491) 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 human Mst2 (Met1-Phe491) (GenBank Accession No. BC010640) 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-Mst2 fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. Mst2 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, 0.6 μM BSA, Substrate: MBP 400 ng/μL and recombinant Mst2: variable.
Quality Control
The theoretical molecular weight of the GST-Mst2 fusion protein is 83 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. Mst2 kinase activity was determined using a radiometric assay [Fig.2].
Background
Mst kinases, members of the STE20 family of kinases, are upstream activators of MAPK pathways that regulate processes such as apoptosis, morphogenesis and cytoskeletal rearrangements. The amino-terminal kinase domain of Mst is considerably homologous to the kinase domain of yeast STE20 kinase and other p21-activated kinases (1). The carboxy-terminal region of Mst1 and Mst2 contains dimerization and inhibitory domains (1-3). Dimerization and phosphorylation at the activation loop results in translocation of Mst1 from the cytosol to the nucleus (3). Growing evidence indicates that Mst1, Mst2 and Mst3 are activated by apoptotic signals as well as other stress conditions (4-6). Complete activation of Mst1 requires both phosphorylation and caspase-mediated cleavage (4). Sequence alignment of the activation loop of the GCK family indicates that Thr183 of Mst1 and Thr180 of Mst2 are the conserved residues and might be critical for the activity of the kinases. Activated Mst kinases may rely on p38 MAPK and JNK pathways to amplify apoptotic signals (5). Phosphorylation at Ser327 of Mst1, which is close to the caspase-3 recognition site, inhibits caspase-mediated cleavage (4).
- Dan, I. et al. (2001) Trends Cell Biol. 11, 220-230.
- Creasy, C.L. et al. (1996) J. Biol. Chem. 271, 21049-21053.
- Lee, K. and Yonehara, S. (2002) J. Biol. Chem. 277, 12351-12358.
- Graves, J.D. et al. (2001) J. Biol. Chem. 276, 14909-14915.
- Lee, K. et al. (2001) J. Biol. Chem. 276, 19276-19285.
- Graves, J.D. et al. (1998) EMBO J. 17, 2224-2234.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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.
