Biological Information
Background Information:
Glycogen synthase kinase-3 (GSK-3) is a protein kinase that was originally identified as a regulator of glycogen synthase, a key enzyme in glycogen metabolism. There are two isoforms of GSK-3, GSK-3a and GSK-3b, and they are strictly regulated via phosphorylation. Phosphorylation of GSK-3b on Ser9 by protein kinase B (PKB) causes its inactivation and is the primary mechanism responsible for growth factor inhibition of this kinase. Activation of GSK-3b is dependent upon the phosphorylation of Tyr216. Upon activation, it has been shown to phosphorylate a number of different cellular proteins, including p53, c-Myc, c-Jun, heat shock factor-1 (HSF-1), and cyclin D1. GSK-3b also has been shown to phosphorylate aberrant sites on the microtubule associated protein tau, which is critical for the progression of Alzheimer’s disease. The inhibitors may have therapeutic opportunities for the treatment of diabetes, Alzheimer's disease, stroke and bipolar disorder.
Family:
Kinase
Sub Family:
CMGC
Class:
Protein Ser/Thr
Protein Name:
GSK3beta
Uniprot Number:
P49841
Protein Aliases:
Serine/threonine-protein kinase GSK3B
Gene Name:
GSK3B
Gene ID:
2932
Accession Number:
NM_001146156
Organism:
Human
Construct Details:
Fragment (2-end), Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Human recombinant protein kinase GSK-3b expressed in Sf21 insect cells is used. Test compound and/or vehicle is preincubated with 500 ng/ml enzyme‡ in modified MOPS buffer pH 7.2 for 15 minutes at 37⁰C. The reaction is initiated by addition of 5 μM phospho-glycogen synthase peptide-2, 10 μM ATP and 0.25 μCi [γ32P]ATP for another 30 minute incubation period and terminated by further addition of 3% H3PO4. An aliquot is removed and counted to determine the amount of [32P]Phospho-glycogen synthase peptide-2 formed. Compounds are screened at 10 μM.
Substrate:
5 µM Phospho-glycogen synthase peptide-2
ATP Concentration:
10 uM
Incubation:
30 min at 37°C
Control Inhibitor:
5-iodo-Indirubin-3'-monoxime
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei