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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 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. Incubate GCK with 0.25 mg/ml Myelin basic Protein (substrate) and 50 ?M [33P]-ATP at 30°C for 15 minutes, then spot reaction on phosphocellulose paper, fix in 1% phosphoric acid, and assay with a scintillation counter.
Background: GCK is a serine/threonine protein kinase that is activated by TNFα and interacts with the TNF receptor-associated factor 2 (TRAF2). GCK is upstream of MAP kinases and particularly activates the SAPK pathway (1). Although GCK is found in many tissues, its expression in lymphoid follicles is restricted to the cells of germinal centre, where it may participate in B-cell differentiation. A mouse protein Rab8ip that has high homology to GCK can interact with the vesicular transport protein Rab8. Based on its interaction with Rab8, it is postulated that Rab8ip/GCK may modulate secretion in response to stress stimuli (2).
Description: Human GCK, also known as MAP4K2, GenBank Accession No. NM_004579, full length, with N-terminal GST tag, MW ~125 kDa, expressed in 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_004579
Host Cell Line: Sf9 cells
Purity: ≥80%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: Q12851
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Pombo C M, et al: Activation of the SAPK pathway by the human STE20 homologue germinal centre kinase. Nature 377: 750-754, 1995.
2. Ren M, et al: In its active form, the GTP-binding protein rab8 interacts with a stress-activated protein kinase. Proc. Nat. Acad. Sci. 93: 5151-5155, 1996.
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