Biological Information

Background Information:

Pepsin produced in the mucosal lining of the stomach acts to degrade protein. Pepsin is one of three principal protein-degrading, or proteolytic, enzymes in the digestive system, the other two being chymotrypsin and trypsin. It is activated by stomach acid. Inhibition of pepsin may be useful to prevent inflammation after gastrointestinal surgery.

Family:

Protease

Sub Family:

Pepsin

Class:

Aspartic Peptidase

Protein Name:

Pepsinogen

Uniprot Number:

P0DJD7

Protein Aliases:

Pepsinogen-4

Gene Name:

PGA4

Gene ID:

643847

Gene Aliases:

pepsin A

Accession Number:

NM_001079808

Organism:

Human

Construct Details:

full length 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 pepsinogen A expressed in mouse myeloma cells is used. Test compound and/or vehicle is preincubated with 10.9 ng/ml enzyme in sodium citrate buffer pH 4.0 for 15 minutes at 25°C. The reaction is initiated by addition of 3 mM MOCAc-Ala-Pro-Ala-Lys-Phe-Phe-Arg-Leu-Lys(Dnp)-NH2 for another 30-minute incubation period. Determination of the amount of MOCAc-Ala-Pro-Ala-Lys-Phe formed is read spectrofluorimetrically at 325 nm/400 nm. Compounds are screened at 10 µM.

Substrate:

3 µM MOCAc-Ala-Pro-Ala-Lys-Phe-Phe-Arg-Leu-Lys(Dnp)-NH2

Incubation:

30 min at 25°C

Control Inhibitor:

Pepstatin

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).

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

5478883

Name

Pepstatin

Measurement

0.00043μM - IC50

Clinical Relevance

Therapeutic Area:

Gastroenterology

Adverse / Beneficial Action:

Inhibition of pepsin may impair protein digestion and assimilation as well as prevent inflammation after gastrointestinal surgery.

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei