Product Pathways - Kinases
KHS1 Kinase #7373
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Description
Purified recombinant full-length human KHS1 (Met1-Tyr846) 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 KHS1 (Met1-Tyr846) (GenBank Accession No. NM_006575) 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 KHS1 protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. KHS1 kinase activity was measured in a radiometric assay using the following reaction conditions: 5 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 0.4 mM EDTA, 5 mM MgCl2, 0.05 mM DTT, 50 μM ATP, Substrate: MBP 200 ng/μL and a variable amount of KHS1.
Quality Control
The theoretical molecular weight of the KHS1 protein is 120 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. KHS1 kinase activity was determined using a radiometric assay [Fig.2].
Background
The KHS1 (?kinase homologous to SPS1/STE20?) serine/threonine protein kinase is a member of the group I germinal center kinase (GCK) family and an upstream component in MAPK signaling. Also referred to as GCKR (?GCK-related kinase,?) KHS1 has an amino-terminal STE20-like protein kinase domain and a carboxyl-terminal regulatory domain that associates with adaptor proteins (1, 2). A widely expressed protein of 95 kDa, KHS1 activates SAPK/JNK in vitro (1). The kinase associates with SH2/SH3 adaptor proteins Crk and CrkL to promote activation of both KHS1 and SAPK/JNK (3). TNF-induced activation of KHS1 can be mediated by TRAF2, which recruits and activates KHS1 resulting in activation of both this kinase and SAPK (4). KHS1 may have important role in regulating cellular stress response (3,4).
- Tung, R.M. and Blenis, J. (1997) Oncogene 14, 653-9.
- Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.
- Shi, C.S. et al. (2000) Blood 95, 776-82.
- Shi, C.S. et al. (1999) J Immunol 163, 3279-85.
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.
