Cysteine-proteinase-inhibiting function of T kininogen and of its proteolytic fragments

Eur J Biochem. 1988 Apr 5;173(1):185-90. doi: 10.1111/j.1432-1033.1988.tb13983.x.

Abstract

Previous attempts to liberate T kinin from T kininogen [Moreau et al. (1986) Eur. J. Biochem. 159, 341-346; Gutman et al. (1988) Eur. J. Biochem. 171, 577-582] have shown that complete fragmentation of the precursor molecule into inhibitory peptides was achieved before any vasoactive peptide was released, suggesting a possible physiological significance for this phenomenon. In this study, cysteine-proteinase-inhibiting properties of rat T kininogen and of its proteolytic fragments issuing from trypsin and submaxillary gland endopeptidase k hydrolysis, have been investigated using rat lysosomal cathepsins B, H and L, papain and bovine calpains I and II. All three lysosomal cathepsins were inhibited by T kininogen but tighter interactions were observed with cathepsin L and papain. Though higher Ki values were obtained for cathepsins B and H, rate constants for association were found to have high and almost similar values (in the 10(6) M-1 s-1 range) whatever the enzyme used. Proteolytic fragments also inhibited cathepsin L and papain very strongly and even better than the entire molecule for some of them, but no significant inhibition of cathepsins B and H was observed. Bovine calpains were not inhibited by T kininogen nor by its proteolytic fragments. From the results of this kinetic analysis, which indicates that both the association and the dissociation of lysosomal cysteine proteinases with T kininogen may occur rapidly, an hypothesis has been put forward on the possible in vivo functioning of T kininogen as a proteinase inhibitor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cathepsin B / pharmacology
  • Cysteine / antagonists & inhibitors*
  • Hydrolysis
  • Kinetics
  • Kininogens / isolation & purification
  • Kininogens / pharmacology*
  • Peptide Fragments / pharmacology*
  • Protease Inhibitors / pharmacology*
  • Rats
  • Substrate Specificity

Substances

  • Kininogens
  • Peptide Fragments
  • Protease Inhibitors
  • Cathepsin B
  • Cysteine