Activity-based probes that target diverse cysteine protease families

Nat Chem Biol. 2005 Jun;1(1):33-8. doi: 10.1038/nchembio707. Epub 2005 May 24.


Proteases are one of the largest and best-characterized families of enzymes in the human proteome. Unfortunately, the understanding of protease function in the context of complex proteolytic cascades remains in its infancy. One major reason for this gap in understanding is the lack of technologies that allow direct assessment of protease activity. We report here an optimized solid-phase synthesis protocol that allows rapid generation of activity-based probes (ABPs) targeting a range of cysteine protease families. These reagents selectively form covalent bonds with the active-site thiol of a cysteine protease, allowing direct biochemical profiling of protease activities in complex proteomes. We present a number of probes containing either a single amino acid or an extended peptide sequence that target caspases, legumains, gingipains and cathepsins. Biochemical studies using these reagents highlight their overall utility and provide insight into the biochemical functions of members of these protease families.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Cysteine Endopeptidases / chemistry*
  • Cysteine Endopeptidases / metabolism
  • Cysteine Proteinase Inhibitors / chemical synthesis
  • Cysteine Proteinase Inhibitors / chemistry*
  • Humans
  • Ketones / chemical synthesis
  • Ketones / chemistry*
  • Molecular Probe Techniques
  • Molecular Probes / chemical synthesis
  • Molecular Probes / chemistry*
  • Substrate Specificity


  • Cysteine Proteinase Inhibitors
  • Ketones
  • Molecular Probes
  • Cysteine Endopeptidases

Associated data

  • PubChem-Substance/840972
  • PubChem-Substance/840973
  • PubChem-Substance/840974
  • PubChem-Substance/840975
  • PubChem-Substance/840976
  • PubChem-Substance/840977
  • PubChem-Substance/840978
  • PubChem-Substance/840979
  • PubChem-Substance/840980
  • PubChem-Substance/840981
  • PubChem-Substance/840982
  • PubChem-Substance/840983
  • PubChem-Substance/840984