Expression, purification, and characterization of caspases

Curr Protoc Protein Sci. 2003 Feb;Chapter 21:21.13.1-21.13.15. doi: 10.1002/0471140864.ps2113s30.

Abstract

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.

MeSH terms

  • Amino Acid Chloromethyl Ketones / metabolism
  • Animals
  • Biological Assay / methods*
  • Caspases* / genetics
  • Caspases* / isolation & purification
  • Caspases* / metabolism
  • Cysteine Proteinase Inhibitors / metabolism
  • Enzyme Activation
  • Humans
  • Isoenzymes* / genetics
  • Isoenzymes* / isolation & purification
  • Isoenzymes* / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Substrate Specificity

Substances

  • Amino Acid Chloromethyl Ketones
  • Cysteine Proteinase Inhibitors
  • Isoenzymes
  • Recombinant Proteins
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • Caspases