Biological Information

Background Information:

Phosphorylase kinase (Phk) is a complex enzyme consisting of four different subunits, (abgd)4, and isoforms or splice variants exist for each subunit. A muscle specific form of Phk deficiency is caused by mutations in the gene for the muscle isoform of the a subunit, PHKA1, whereas liver Phk deficiency can be caused by mutations in three genes: PHKA2, PHKB, and PHKG2. PHKB mutations cause autosomal recessive Phk deficiency of both liver and muscle, but liver symptoms predominate and the biochemical muscle involvement is often not clinically apparent. Ten different mutations in the PHKB gene have been identified in seven patients. PHKG2, also autosomal, encodes the testis/liver isoform of the catalytic g subunit. PHKG2 is a serine/threonine kinase that plays an important role in glycogen metabolism. Inhibition of PHKG2 may result in recurrent hypoglycemia, liver fibrosis and increased risk of cirrhosis.

Family:

Kinase

Sub Family:

CAMK

Class:

Protein Ser/Thr

Protein Name:

PHKG2

Uniprot Number:

P15735

Protein Aliases:

PHK-gamma-T|PSK-C3|Phosphorylase kinase subunit gamma-2

Gene Name:

PHKG2

Gene ID:

5261

Accession Number:

NM_000294

Organism:

Human

Construct Details:

Full Length, 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 PHKG2 expressed in insect Sf9 cells is used. Test compound and/or vehicle is preincubated with 0.364 μg/ml enzyme‡ in modified MOPS buffer pH 7.2 for 15 minutes at 37⁰C. The reaction is initiated by addition of 50 μM ZIPtide, 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]ZIPtide formed. Compounds are screened at 10 μM.

Substrate:

50 µM ZIPtide

ATP Concentration:

10 uM

Incubation:

30 min at 37°C

Control Inhibitor:

Staurosporine

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

PubChem ID

122130844

Name

Staurosporine

Measurement

0.0018μM - IC50

Clinical Relevance

Therapeutic Area:

Metabolic Diseases

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

RELATED SOLUTIONS

Item: PP90