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

PubChem ID

122130844

Name

Staurosporine

Measurement

2.5nM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

KP - France - Celle L'Evescault

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Item: 14-608KP