Apoptotic caspase activation and activity.
Caspases are central to the execution of apoptosis. Their proteolytic activity is responsible for the demise of cells in many physiological and pathological states. Great advances in understanding caspases have been made using recombinant caspase expression and enzymatic characterization. Assays to measure caspase activity in apoptotic cell extracts and the development of a reconstituted cell-free assay were also critical in establishing the hierarchy in the caspase activation cascade and comprehend how caspase-9 is activated by the apoptosome. More recently, new tools such as activity-based probes allowed us to detect caspase activation in their working environment providing readout of the system with minimal interference. This chapter describes some of the methods used by our group to study the activation mechanisms of caspases and their activity.
Denault
Jean-Bernard
JB
The Burnham Institute for Medical Research, La Jolla, CA, USA.
Salvesen
Guy S
GS
D017209:Apoptosis / Q000502:physiology*; D020169:Caspases / Q000378:metabolism*; D002457:Cell Extracts / Q000032:analysis; D004789:Enzyme Activation; D006801:Humans; D015151:Immunoblotting; D011994:Recombinant Proteins / Q000378:metabolism; D013194:Staining and Labeling; D019809:Streptavidin / Q000032:analysis
D002457:Cell Extracts; D011994:Recombinant Proteins; D019809:Streptavidin; D020169:Caspases