Biological Information

Background Information:

Caspases are the executioners of programmed cell death (apoptosis), a normal process used physiologically to eliminate specific cells or cell types without damaging surrounding cells or causing an inflammatory response. All caspases belong to a family of proteases characterized by a cysteine residue in the active site and display specificity for Asp residues. Caspases are present in all cells as a procaspase, which becomes activated through proteolytic cleavage by other proteases including other caspases. Caspase-1 (interleukin-1 beta-converting enzyme, ICE) is responsible for proteolytic activation of the biologically inactive interleukin-1 beta precursor to the proinflammatory cytokine. Inhibition of ICE is a new anti-inflammatory strategy being explored by the design of both reversible inhibitors and irreversible inactivators of the enzyme. ICE inhibitors that cross react against multiple ICE homologs can also block diverse types of apoptosis.

Family:

Protease

Sub Family:

Caspase

Class:

Cysteine Peptidase

Protein Name:

Caspase 1

Uniprot Number:

P29466

Protein Aliases:

caspase-1|interleukin 1, beta, convertase

Gene Name:

CASP1

Gene ID:

834

Gene Aliases:

ICE

Accession Number:

NM_033292

Organism:

Human

Construct Details:

Fragment, 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 Caspase 1 expressed in E. coli cells is used. Test compound and/or vehicle are preincubated with 2.5 U/ml‡ of enzyme for 15 minutes at 30⁰C in Hepes buffer pH 7.4. The reaction is initiated by addition of 7.5 µM Z-YUAD-AFC for another 45 minute incubation period. Read plate on a spectrophotometer (Excitation: 405 nm, Emission: 535 nm). Compounds are screened at 10 μM.

Substrate:

7.5 µM Z-YUAD-AFC

Incubation:

45 min at 30°C

Control Inhibitor:

Ac-YVAD-CHO

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

644345

Name

Ac-DEVD-CHO

Measurement

0.046μM - IC50

PubChem ID

5311139

Name

Ac-YVAD-CHO

Measurement

0.007μM - IC50

PubChem ID

10256708

Name

Ac-LEHD-CHO

Measurement

0.071μM - IC50

PubChem ID

44135117

Name

Ac-IETD-CHO

Measurement

0.19μM - IC50

Clinical Relevance

Therapeutic Area:

Inflammation/Allergy

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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