Biological Information

Background Information:

Ectonucleotide pyrophosphatase/phosphodiesterase family member 2 (ENPP 2), also known as autotaxin, is produced as a pre-proenzyme glycoprotein and activated by the removal of its N-terminal signal peptide resulting in a matured secreted form, consists of 4 domains, two N‐terminal SMB‐like domains, a central catalytic PDE domain, and an inactive nuclease‐like domain. ENPP2 acts as a lysophospholipase D, mainly hydrolyzing extracellular lysophosphatidylcholine (LPC) to lysophosphatidic acid (LPA). LPA, like a growth factor in serum, could induce motility in fibroblasts and cancer cells through GPCRs. Deregulation of the LPA signaling axis has been linked to different diseases, such as liver disease, fibrosis, pruritus, multiple sclerosis, inflammation, and cancer. A specific ENPP2 inhibitor against idiopathic pulmonary fibrosis showed promising results in the clinical trial, and the inhibition of ENPP2 also can stimulate T-cell infiltration into tumors and enhance the efficacy of immunotherapy for liver disease and pancreatic cancer.

Family:

Phosphatase

Class:

Protein (enzyme)

Protein Name:

ENPP2

Uniprot Number:

Q13822

Protein Aliases:

Autotaxin; Extracellular lysophospholipase D (LysoPLD)

Gene Name:

ENPP2

Gene ID:

5168

Gene Aliases:

ATX; PD-IALPHA

Accession Number:

NM_001040092

Organism:

Human

Construct Details:

Fragment, Recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Spectrofluorimetry

Platform:

Plate reader

Measured Response:

Fluorescence

Testing Information

Procedure Summary:

Human recombinant protein ENPP2 expressed in mouse myeloma cells is used. Test compound and/or vehicle is preincubated with 0.136 µg/ml enzyme for 15 minutes at 37°C in modified Tris-HCl buffer pH 8.0. The reaction is initiated by addition of 0.5 µM FS-3 for 30 minutes. Determination of the amount of divided fluorescence parts formed is read spectrofluorimetrically at 485 nm/520 nm. Compounds are screened at 10 µM.

Substrate:

0.5 µM FS-3

Incubation:

30 min at 37°C

Control Inhibitor:

ONO-8430506

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

29µM - IC50

PubChem ID

56597976 

Name

ONO-8430506

Measurement

0.00041µM - IC50

PubChem ID

46911532

Name

HA 130

Measurement

0.0033µM - IC50

PubChem ID

25265312

Name

PF 8380

Measurement

0.00051µM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei