Cysteine-proteinase-inhibiting function of T kininogen and of its proteolytic fragments.
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.
Moreau
T
T
Laboratoire de Biochimie, Faculté de Médecine, Université François Rabelais, Tours, France.
Esnard
F
F
Gutman
N
N
Degand
P
P
Gauthier
F
F
D000595:Amino Acid Sequence; D000818:Animals; D002401:Cathepsin B / Q000494:pharmacology; D003545:Cysteine / Q000037:antagonists & inhibitors*; D006868:Hydrolysis; D007700:Kinetics; D007704:Kininogens / Q000302:isolation & purification / Q000494:pharmacology*; D010446:Peptide Fragments / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000494:pharmacology*; D051381:Rats; D013379:Substrate Specificity
D007704:Kininogens; D010446:Peptide Fragments; D011480:Protease Inhibitors; D002401:Cathepsin B; D003545:Cysteine