Biological Information
Background Information:
Death-associated protein kinase (DAPK) is a calcium/calmodulin-dependent kinase that phosphorylates serine/threonine residues on proteins. DAPK modulates cell death induced by IFN-g, TNF-a, Fas, and detachment from the extracellular matrix. DAPK has been also implicated in TGF-binduced apoptosis in several cultured cell lines in which Smad proteins mediate transcriptional activation of DAPK. DAPK3, also known as Zipper interacting protein (ZIP) kinase, contains an N-terminal kinase domain and a C-terminal leucine zipperstructure and binds to ATF4 that is a member of ATF/CREB family. Overexpression of ZIP kinase induces apoptosis. ZIP has also been shown to have a potential role in regulating VSMC contraction.
Family:
Kinase
Sub Family:
CAMK
Class:
Protein Ser/Thr
Protein Name:
DAPK3 (ZIP)
Uniprot Number:
O43293
Protein Aliases:
DAP-like kinase|Dlk|MYPT1 kinase|Zipper-interacting protein kinase|ZIP-kinase
Gene Name:
DAPK3
Gene ID:
1613
Gene Aliases:
ZIP|ZIPK
Accession Number:
AB007144
Organism:
Human
Construct Details:
1-290 recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometric
Measured Response:
Scintillation
Testing Information
Procedure Summary:
ZIPK (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 250 µM KKLNRTLSFAEPG, 10 mM MgAcetate and [g- 33P]-ATP (specific activity and concentration as required). The reaction is initiated by the addition of the Mg/ATP mix. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the reaction is then spotted onto a filter and washed four times for 4 minutes in 0.425% phosphoric acid and once in methanol prior to drying and scintillation counting.
Substrate:
250 uM KKLNRTLSFAEPG
Tracer:
33P
ATP Concentration:
[ATP]= 10µM (Km= 14µM )
Incubation:
40 min at Room temperature
Control Inhibitor:
Staurosporine
Test Sample Requirements:
Please provide compounds dissolved in 100% DMSO (PBS or HBSS prohibited).
Stock solution should be ≥50× the final assay concentration (a 10 mM stock is also acceptable).
Required volume:
– <60 kinase assays: 120 µL of 50× stock
– ≥60 assays: 420 µL of 50× stock
If you supply 10 mM stocks or powder (pre-weighed), we will apply the appropriate conversion.
For batches of ~50 compounds supplied as solids, the TAT will be extended.
Minimum Order Quantity:
2
Turnaround Time
Standard:
6 Business Days
Service Scheduling Note:
Assay is run on a weekly schedule beginning on Friday. Compounds and purchase order must be received on or before 13:00 CET Thursday for the order to be included on the service run that begins on Friday of the same week.
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Additional Information
Brand:
LeadHunter
Testing Location:
KP - France - Celle L'Evescault
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