beta-Secretase 1 Human Pepsin Aspartic Peptidase Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
The amyloid beta peptides (Aß) are the major components of the senile plaques characteristic of Alzheimer's disease. Aß peptides are generated from the cleavage of amyloid precursor protein (APP) by beta- and gamma-secretases. beta-Secretase (BACE1, BACE, beta-site APP-cleaving enzyme), a type-I transmembrane aspartyl protease, cleaves APP first to generate a 99-amino acid membrane-associated fragment (CT99) containing the N terminus of Aß peptides. beta-Secretase then cleaves within the transmembrane region of CT99 to generate the C terminal of Aß peptides. The cleavage of APP by BACE1 is believed to be a prerequisite for gamma-secretase-mediated processing. Inhibition of BACE1 may be of therapeutic benefit in the treatment of Alzheimer's disease.
Family:
Protease
Sub Family:
Pepsin
Class:
Aspartic Peptidase
Protein Name:
beta-Secretase 1
Uniprot Number:
P56817
Protein Aliases:
Aspartyl protease 2; Beta-site amyloid precursor protein cleaving enzyme 1; Memapsin-2; Membrane-associated aspartic protease 2
Gene Name:
BACE1
Gene ID:
23621
Gene Aliases:
BACE
Accession Number:
NM_001207048
Organism:
Human
Construct Details:
Fragment (22-460), Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human recombinant beta-secretase expressed in murine myeloma NS0 cells is used. Test compound and/or vehicle is preincubated with 8 µg/ml enzyme in acetate buffer pH 4.0 for 15 minutes at 37˚C. The reaction is initiated by addition of 10 µM MOCAc-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Arg-Lys(Dnp)-Arg-Arg-NH2 for another 60-minute incubation period. Determination of the amount of MOCAc-Ser-Glu-Val-Asn-Leu formed is read spectrofluorimetrically with excitation at 328 nm and emission at 393 nm. Compounds are screened at 10 µM.
Substrate:
10 µM MOCAc-S-E-V-N-L-D-A-E-F-R-K(Dnp)-R-R-NH2
Incubation:
60 min at 37°C
Control Inhibitor:
beta-Secretase Inhibitor IV
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:
CNS/Addiction/Pain
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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