Expression, purification, and characterization of caspases.
This unit describes a protocol to obtain milligram amounts of enzymatically active pure recombinant caspases. Specific details for the expression, purification of caspase-3, -6, -7, -8, -9 and -10 are discussed along with strategies to obtain particular forms (e.g., the zymogen) of some of them.
Denault
Jean-Bernard
JB
The Burnham Institute, La Jolla, California.
Salvesen
Guy S
GS
The Burnham Institute, La Jolla, California.
D000590:Amino Acid Chloromethyl Ketones / Q000378:metabolism; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D020169:Caspases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D004789:Enzyme Activation; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
D000590:Amino Acid Chloromethyl Ketones; D015853:Cysteine Proteinase Inhibitors; D007527:Isoenzymes; D011994:Recombinant Proteins; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D020169:Caspases