Biological Information
Background Information:
Ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), a metalloenzyme, is a homodimer type II transmembrane glycoprotein characterized by an N-terminal transmembrane domain, two somatomedin-B-like domains, a catalytic domain, and a C-terminal nuclease-like domain. ENPP1 hydrolyzes a wide range of phosphodiester bonds. The main substrate of ENPP1 is ATP, which is cleaved into AMP and diphosphate. Another notable nucleotide substrate is NAD+ which can be hydrolyzed to produce AMP. The enzyme is involved in bone mineralization, soft-tissue calcification, insulin receptor signaling, cancer cell proliferation, and immune modulation. Diseases associated with ENPP1 include generalized arterial calcification of infancy, ossification of the posterior longitudinal ligament of the spine (OPLL), hypophosphatemic rickets autosomal recessive 2 (ARHR2), and insulin resistance.
Family:
Phosphatase
Class:
Protein (enzyme)
Protein Name:
ENPP1
Uniprot Number:
P22413
Protein Aliases:
Membrane component chromosome 6 surface marker 1
Gene Name:
ENPP1
Gene ID:
5167
Gene Aliases:
PC-1; PCA1
Accession Number:
NM_006208
Organism:
Human
Construct Details:
Fragment, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrophotometry
Platform:
Plate reader
Measured Response:
Absorbance
Testing Information
Procedure Summary:
Human recombinant protein ENPP1 expressed in HEK293 cells is used. Test compound and/or vehicle is preincubated with 5.34 U/ml enzyme for 15 minutes at 25°C in modified Tris-HCl buffer pH 9.5. The reaction is initiated by addition of 0.3 mM Thymidine 5’-monophosphate p-nitrophenyl ester for 30 minutes. Determination of the amount of 4-Nitrophenol formed is read spectrophotometrically at 405 nm. Compounds are screened at 10 µM.
Substrate:
300 µM Thymidine 5’-monophosphate p-nitrophenyl ester
Incubation:
30 min at 25°C
Control Inhibitor:
ENPP1-IN-1
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