Dynorphin-derived peptides reveal the presence of a critical cysteine for the activity of brain endo-oligopeptidase A

Biochem Biophys Res Commun. 1993 Dec 15;197(2):501-7. doi: 10.1006/bbrc.1993.2507.

Abstract

Brain endo-oligopeptidase A, a neuropeptide-metabolizing endopeptidase, has been considered a cysteine-endopeptidase because it is activated by thiols and inhibited by phydroxymercuribenzoate or 5,5'-dithiobis-(2-nitrobenzoic acid). The understanding of the unique specificity of endo-oligopeptidase A was useful for the synthesis of affinity labeling compounds containing as a thiol reactive group the Cys-(3-nitro-2-pyridinesulfenyl) group into dynorphin-derived peptides which are among the best substrates and competitive inhibitor of endopeptidase 22.19. Of the ten compounds tested, only peptides containing 8 to 13 amino acid residues caused irreversible inhibition. The fact that the most effective inhibitors had the reactive group either at the P'1 or at P'3 position [nomenclature of Schechter and Berger] would seem to argue that the reactive cysteine is in the vicinity of the active site, or actually involved in the catalytic step.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Brain / enzymology*
  • Cattle
  • Chromatography, High Pressure Liquid
  • Cysteine*
  • Dynorphins / chemistry*
  • Dynorphins / metabolism
  • Kinetics
  • Metalloendopeptidases / isolation & purification
  • Metalloendopeptidases / metabolism*
  • Molecular Sequence Data
  • Oligopeptides / chemical synthesis
  • Oligopeptides / pharmacology*
  • Peptide Fragments / chemistry*
  • Peptide Fragments / isolation & purification
  • Peptide Fragments / metabolism
  • Substrate Specificity

Substances

  • Oligopeptides
  • Peptide Fragments
  • Dynorphins
  • Metalloendopeptidases
  • thimet oligopeptidase
  • Cysteine