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Encompassing Amino Acids: 156-555
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 HIPK1 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: HIPK1 or homeodomain-interacting protein kinase 1 is a ser/thr protein kinase and a member of the HIPK family. HIPK 1 is a nuclear kinase that phosphorylates homeodomain transcription factors. HIPK1 phosphorylates DAXX and this leads to its relocalization and subsequent decrease in transcriptional repression activity (1). HIPK1also interacts with p53 and phosphorylates it on serine residues. HIPK 1 expression is elevated in breast cancer cell lines and embryonic fibroblasts from HIPK 1-null mice show more susceptibility to apoptosis induced by DNA damage (2).
Description: Human HIPK1 (homeodomain-interacting protein kinase 1), also known as Myak and Nbak2 (GenBank Accession No. NM_152696), a.a. 156-555 with N-terminal GST tag, MW = 71 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_152696
Host Cell Line: Sf9 cells
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: Q86Z02
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Ecsedy, J A. et al: Homeodomain-interacting protein kinase 1 modulates Daxx localization, phosphorylation, and transcriptional activity. Mol Cell Biol. 2003 Feb;23(3):950-60.
2. Kondo, S. et al: Characterization of cells and gene-targeted mice deficient for the p53-binding kinase homeodomain-interacting protein kinase 1 (HIPK1). Proc Natl Acad Sci U S A. 2003 Apr 29;100(9):5431-6.
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