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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: The assay is performed in 5 mM MOPS, 2.5 mM β-glycerophosphate, 5 mM MgCl2, 1 mM EGTA, 0.4 mM EDTA, 0.05 mM DTT, 2 mM ATP. MSK1 is incubated with a short peptide substrate (KRRRLSSLRA) and 50 uM [33P]-ATP at 30°C for 15 minutes, then the reaction is spotted onto phosphocellulose paper, fixed in 1% phosphoric acid, and assayed with a scintillation counter.
Background: MSK1 or mitogen- and stress-activated protein kinase-1 contains 2 protein kinase domains and shares 43% protein sequence identity with the MAPKAPK1 isoforms. Northern blot analysis shows that MSK1 is expressed in all tissues with the highest level of expression in brain, muscle, and placenta. Immunoelectron microscopy localized MSK1 to the nucleus. MSK1 is activated in vitro and in vivo by either ERK or SAPK2 proteins. MSK1 rather than MAPKAP-K1 or MAPKAP-K2/K3, mediates activation of the cAMP response element-binding protein and activating transcription factor-1 by either growth factors or stress signals. By radiation hybrid analysis, the RPS6KA5 gene has been mapped to chromosome 14q31-q32.
Description: Recombinant human MSK1, full length, with N-terminal GST-tag, expressed by Sf9 insect cells via a baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.25 mM DTT, 0.1 mM EGTA, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol.
Genbank: NM_004755
Host Cell Line: Sf9 cells
Purity: ≥75%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: O75582
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Deak, M. et al: Mitogen- and stress-activated protein kinase-1 (MSK1) is directly activated by MAPK and SAPK2/p38, and may mediate activation of CREB. EMBO J. 17: 4426-4441, 1998.
2. Jiang, C. et al: Assignment of a member of the ribosomal protein S6 kinase family, RPS6KA5, to human chromosome 14q31-q32.1 by radiation hybrid mapping. Cytogenet. Cell Genet. 87: 261-261, 1999.
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