Product Pathways - MAPK Signaling
RSK3 Kinase #7408
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 RSK3 (Met1-Leu733) 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 RSK3 (Met1-Leu740) (GenBank Accession No. NM_021135) with an amino-terminal GST tag. The protein was purified by one-step affinity chromatography using GSH-agarose.
Gel Staining
Figure 1. The purity of the GST-RSK3 fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. RSK3 kinase activity was measured in a radiometric assay using the following reaction conditions: 5 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 5 mM EGTA, 0.4 mM EDTA, 5 mM MgCl2, 0.05 mM DTT, 50 μM ATP, Substrate: S6K synthetic peptide substrate, 400 ng/μL, and recombinant RSK3: variable.
Quality Control
The theoretical molecular weight of the GST-RSK3 fusion protein is 112 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. RSK3 kinase activity was determined using a radiometric assay [Fig.2].
Background
The 90 kDa ribosomal S6 kinases (RSK1?4) are a family of widely expressed serine/threonine kinases characterized by two nonidentical, functional kinase domains (1) and a C-terminal docking site for extracellular signal-regulated kinases (ERKs) (2). Several sites both within and outside of the RSK kinase domain, including Ser380, Thr359, Ser363 and Thr573, are important for kinase activation (3). RSK1-3 are activated via coordinated phosphorylation by MAPKs, by autophosphorylation, and by phosphoinositide-3-OH kinase (PI3K) in response to many growth factors, polypeptide hormones and neurotransmitters (3).
- Fisher, T.L. and Blenis, J. (1996) Mol Cell Biol 16, 1212-9.
- Smith, J.A. et al. (1999) J Biol Chem 274, 2893-8.
- Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
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.
