Biological Information

Background Information:

Glucokinase catalyzes the formation of glucose-6-phosphate, expressed in the pancreatic beta cell and liver involved in sensing systemic glucose and regulating glucose homeostasis.

Family:

Kinase

Protein Name:

GCK

Uniprot Number:

P35557 

Protein Aliases:

Glucokinase 

Accession Number:

NM_000162

Organism:

Human

Construct Details:

Fragment (16-465), Recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Agonist

Detection Method:

Spectrophotometry

Platform:

Spectrofluorimetry

Testing Information

Procedure Summary:

Human recombinant Glucokinase (GCK) expressed in E. coli cells is used. Test compound or 100 µM PF 04991532 or vehicle is preincubated with 4 µg/ml enzyme in modified HEPES buffer pH 7.0 for 15 minutes at 25C. The reaction is initiated by addition of 20 mM Glucose, 200 µM ATP and 2 µg/ml Coupling Phosphatase 4 for another 20 minute incubation period. The amount of inorganic phosphate are determined spectrophotometrically at 620 nm by malachite green reagent. Compounds are screened at 10 µM.

Substrate:

20 µM Glucose

Incubation:

20 min at 25°C

Control Inhibitor:

PF 04991532

Criteria For Significance:

≥ 50% of max stimulation

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:

10

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

46181428

Name

PF 04991532

Measurement

0.54μM - IC50

Clinical Relevance

Therapeutic Area:

Metabolic Diseases

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei