Regulatory Status Legend

Description

Members of the ICE/CED-3 cysteine protease family have key roles in inflammation and mammalian apoptosis. The ICE family member Caspase-3 (also known as CPP32, Yama, apopain) is activated early in apoptosis and appears to be involved in the proteolysis of several important molecules, including poly (ADP ribose) polymerase (PARP). Activated Caspase-3 cleaves PARP from its 116 kDa to an 85 kDa residual fragment. The cleavage site in PARP is C-terminal to Asp-216. The upstream sequence of the cleavage site, DEVD (Asp-Glu-Val-Asp), is utilized as a basis for the highly specific Caspase-3 substrate, Ac (N-acetyl)-DEVD-AMC (7-amino-4-methylcoumarin).

Ac-DEVD-AMC is a synthetic tetrapeptide fluorogenic substrate for Caspase-3 (CPP32) and contains the amino acid sequence of the PARP cleavage site at Asp-216. The tetrapeptide substrate can be used to identify and quantify the Caspase-3 activity in apoptotic cells. Caspase-3 cleaves the tetrapeptide between D and AMC, thus releasing the fluorogenic AMC, which can be quantified in a spectrofluorometer. The substrate can also be used to study the inhibition of Caspase-3 by the tetrapeptide aldehyde, Ac-DEVD-CHO or any other inhibitor of Caspase-3.