Biological Information

Background Information:

Cathepsin D is a major intracellular member of the mammalian aspartic proteinase family. The enzyme is thought to be in the normal degradation of intracellular and extracellular proteins. It has also been suggested that the enzyme plays a role in the generation of biologically active peptides, the processing of exogenous antigens and the b-amyloid precursor protein in Alzheimer’s disease. Overexpression of cathepsin D in human breast cancers is associated with higher risk of relapse and metastasis. Moreover, recent interest in this enzyme arose from the discovery of its possible role in the execution of cell death and the progression of neoplastic diseases.

Family:

Protease

Sub Family:

Pepsin

Class:

Aspartic Peptidase

Protein Name:

Cathepsin D

Uniprot Number:

P07339

Protein Aliases:

ceroid-lipofuscinosis, neuronal 10

Gene Name:

CTSD

Gene ID:

1509

Gene Aliases:

CLN10

Accession Number:

NM_001909

Organism:

Human

Construct Details:

Endogenous

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Spectrofluorimetry

Measured Response:

Fluorescence

Testing Information

Procedure Summary:

Human liver cathepsin D is used. Test compound or vehicle is preincubated with 190 mU/ml enzyme‡ in modified acetate buffer pH 4.0 for 15 minutes at 37°C. The reaction is initiated by addition of 20 μM MOCAc-Gly-Lys-Pro-Ile-Leu-Phe-Phe-Arg-Leu-Lys(Dnp)-D-Arg NH2 for another 10 minute incubation period. Determination of the amount of MOCAc formed is read spectrofluorimetrically at 340 nm/400. Compounds are screened at 10 μM.

Substrate:

20 µM MOCAc-Gly-Lys-Pro-Ile-Leu-Phe-Phe-Arg-Leu-Lys(Dnp)-D-Arg-NH₂

Incubation:

10 min at 37°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).

Minimum Order Quantity:

2

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

173830

Name

L-Leu-Chloromethylketone

Measurement

>100μM - IC50

PubChem ID

299927

Name

Z-Gly-Leu-Phe-CMK

Measurement

>100μM - IC50

PubChem ID

5478883

Name

Pepstatin

Measurement

0.00093μM - IC50

PubChem ID

7098622

Name

Leupeptin

Measurement

>100μM - IC50

PubChem ID

11957542

Name

E-64

Measurement

>100μM - IC50

PubChem ID

16760356

Name

Cathepsin Inhibitor I

Measurement

>100μM - IC50

PubChem ID

16760357

Name

Cathepsin Inhibitor II

Measurement

>100μM - IC50

PubChem ID

46936189

Name

Chymostatin

Measurement

>100μM - IC50

PubChem ID

Name

Gly-Phe-CHN2

Measurement

>100μM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

RELATED SOLUTIONS

Item: PP90
Item: PP326