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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 HIPK4 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: HIPK4 or homeodomain interacting protein kinase 4 is a serine/threonine protein kinase that can act as a co-repressor of transcription factors (1). HIPK4 has been shown to phosphorylate the human p53 protein at serine 9, both in vitro and in vivo which is important for p53 mediated transcriptional repression. RT-PCR analysis showed moderate expression of HIPK4 in lung and white adipose tissue and weak expression in brain and liver. HIPK4 expression has also been observed in various cancer cell lines (2).
Description: Human HIPK4 (homeodomain-interacting protein kinase 4), GenBank Accession No. NM_144685, full length, with N-terminal GST tag, MW = 110 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_144685
Host Cell Line: Sf9 cells
Purity: ≥70%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: Q8NE63
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Arai, S. et al: Novel homeodomain-interacting protein kinase family member, HIPK4, phosphorylates human p53 at serine 9. FEBS Lett. 2007;11;581(29):5649-57.
2. Di Stefano, V. et al: Homeodomain-interacting protein kinase-2 activity and p53 phosphorylation are critical events for cisplatin-mediated apoptosis. Exp. Cell. Res. 2004;15;293(2):311-20.
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