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

PubChem ID

137524159 

Name

ENPP1-IN-1

Measurement

0.0014µM - IC50

PubChem ID

56597976 

Name

ONO-8430506

Measurement

>100µM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei