Affinity purification of endothia protease with a novel renin inhibitor SQ 32,970

Biochem Biophys Res Commun. 1989 May 30;161(1):1-7. doi: 10.1016/0006-291x(89)91551-9.

Abstract

A novel tripeptidic renin inhibitor is described, SQ 32,970, that will potently inhibit endothia protease. This inhibitor can be coupled to Sepharose and will allow the affinity-purification of endothia protease in one step to greater than 95% purity as measured by SDS PAGE. The purified endothia protease cleaves the Lys-Pro-Ala-Glu-Phe-Nph-Arg-Leu substrate at the Phe-Nph bond with a Kcat/Km of 7445 (s-1 mM-1) at pH 3.1 and 4057 (s-1 mM-1) at pH 6.0. Affinity purified endothia protease can be crystallized in the pH range in which it is enzymatically active and can be inhibited by renin inhibitors.

MeSH terms

  • Ascomycota / analysis*
  • Aspartic Acid Endopeptidases*
  • Chromatography, High Pressure Liquid
  • Endopeptidases / isolation & purification*
  • Fungal Proteins / isolation & purification*
  • Hydrolysis
  • Kinetics
  • Molecular Weight
  • Oligopeptides / pharmacology*
  • Protease Inhibitors
  • Renin / antagonists & inhibitors*
  • Substrate Specificity

Substances

  • Fungal Proteins
  • Oligopeptides
  • Protease Inhibitors
  • SQ 32970
  • Endopeptidases
  • Aspartic Acid Endopeptidases
  • Endothia aspartic proteinase
  • Renin