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Updated: May 12, 2026
05:29
Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry
Published on: March 24, 2023
Caspase functions in cell death and disease.
David R McIlwain1, Thorsten Berger, Tak W Mak
1The Campbell Family Institute for Breast Cancer Research and Ontario Cancer Institute, University Health Network, Toronto, Ontario M5G 2C1, Canada.
Cold Spring Harbor Perspectives in Biology
|April 3, 2013
Summary
Caspases are enzymes crucial for cell death and inflammation. Understanding their controlled activation is key to developing therapies for diseases like cancer and inflammatory disorders.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Caspases are critical endoproteases regulating cell death and inflammation.
- Their activation involves conversion from inactive zymogens to active forms through aggregation.
- Dysregulation of caspases is implicated in cancer and inflammatory diseases.
Purpose of the Study:
- To elucidate the activation mechanisms of caspases.
- To understand the role of caspases in cellular regulatory networks.
- To provide insights for developing targeted therapies for caspase-related diseases.
Main Methods:
- The study focuses on the biochemical and cellular mechanisms of caspase activation.
- Analysis of signaling events leading to caspase aggregation and activity.
- Investigating substrate interactions and downstream effects of caspase activation.
Main Results:
- Caspase activation is tightly regulated by aggregation into dimers or macromolecular complexes.
- Apoptotic caspases execute controlled cellular demolition.
- Inflammatory caspases drive innate immune responses via proinflammatory cytokines.
Conclusions:
- Caspase activity is essential for both apoptosis and inflammation.
- Understanding caspase regulation is vital for treating diseases linked to their dysregulation.
- Targeting caspase pathways offers therapeutic potential for cancer and inflammatory disorders.
Related Concept Videos
01:24Caspases
Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside cells.
01:30Overview of Cell Death
Cell death is an essential process where the body gets rid of old or damaged cells. Cell proliferation and death need to be balanced, as an imbalance between the two may lead to cancer or autoimmune diseases.
Cell death was observed in the early 19th century, but there was no experimental evidence to prove it. In 1842, Carl Vogt first discovered cell death in a metamorphic toad; however, it was not termed ‘cell death.’ Scientists discovered different cell death pathways only in the 20th century...
Cell death was observed in the early 19th century, but there was no experimental evidence to prove it. In 1842, Carl Vogt first discovered cell death in a metamorphic toad; however, it was not termed ‘cell death.’ Scientists discovered different cell death pathways only in the 20th century...
01:17The Extrinsic Apoptotic Pathway
The extrinsic apoptotic pathway is initiated when extracellular death-inducing signals, such as specific cytokines, activate the death receptors expressed on the cell surface. The immune cells involved in this pathway are natural killer cells (NK cells) and cytotoxic T-lymphocytes. NK cells are critical in innate immune response, while cytotoxic T-lymphocytes are associated with adaptive immune response. These cells recognize specific receptors expressed on the altered cells and activate...
01:31The Intrinsic Apoptotic Pathway
Internal cellular stress, such as cellular injury or hypoxia, triggers intrinsic apoptosis. The B-cell lymphoma 2 (Bcl-2) family of proteins are the primary regulators of the intrinsic apoptotic pathway. For example, during DNA damage, checkpoint proteins, such as Ataxia Telangiectasia Mutated (ATM protein) and Checkpoints Factor-2 (Chk2) proteins, are activated. These proteins phosphorylate p53 which further activates pro-apoptotic proteins, such as Bax, Bak, PUMA, and Noxa, and inhibits...
01:30Apoptosis
Apoptosis is a combination of two Greek words, 'apo' and 'ptosis,' meaning separation and falling off, respectively. Hippocrates used this word to describe gangrene, which was caused due to bandaging of fractured bones. Apoptosis was distinguished from necrosis in 1970 when John Kerr reported observations of morphological changes occurring during apoptosis. During one experiment, he observed that the disruption of blood supply to the liver tissue resulted in a size reduction of the tissue.
01:22Cellular Injury V: Apoptosis and Autophagy
Cells respond to damage and stress through highly coordinated processes that decide whether they survive or undergo controlled self-destruction. Two major pathways involved in this regulation are apoptosis, a type of programmed cell death, and autophagy, a survival mechanism that helps cells adapt to adverse conditions.ApoptosisApoptosis removes aged or injured cells to maintain tissue balance. During this process, the cell shrinks, chromatin condenses and fragments, and membrane-bound...