title: Inhibition of tissue kallikrein by protein C inhibitor. Evidence for identity of protein C inhibitor with the kallikrein binding protein.
abstract: We studied the inhibition of tissue kallikrein by protein C inhibitor (PCI), a relatively unspecific heparin-dependent serine protease inhibitor present in plasma and urine. PCI inhibited the amidolytic activity (cleavage of H-D-valyl-L-leucyl-arginine-p-nitroaniline) of urinary kallikrein with an apparent second order rate constant of 2.3 x 10(4) M-1 s-1 and formed stable complexes (85 kDa) with urinary kallikrein as judged from silver-stained sodium dodecyl sulfate-polyacrylamide gels. Complex formation was time-dependent and was paralleled by a decrease in the intensity of the main PCI protein band (Mr = 57,000) and an increase in the intensity of the lower Mr (54,000) PCI form (cleaved inhibitor). Heparin interfered with the inhibition of tissue kallikrein by PCI and with the formation of tissue kallikrein-PCI complexes in a dose-dependent fashion and completely abolished PCI-tissue kallikrein interaction at 300 micrograms/ml. This is in contrast to findings on the interaction of PCI with all other target proteases studied so far (i.e. stimulation of inhibition by heparin) but is similar to the reaction pattern of 125I-labeled tissue kallikrein with so called kallikrein binding protein described in serum and other systems. To study a possible relationship between PCI and this kallikrein binding protein we incubated 125I-labeled urinary kallikrein in serum and in PCI-immunodepleted serum in the absence and presence of heparin and analyzed complex formation using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In normal serum, formed complexes co-migrated with complexes of purified PCI and 125I-kallikrein and were less intense in the presence of heparin. No complex formation at all was seen in PCI-depleted serum. Our data indicate that PCI may be a physiologically important endogenous inhibitor of tissue kallikrein and provide evidence that PCI may be identical to the previously described kallikrein binding protein.
authors.affiliation: Department of Medical Physiology, University of Vienna, Austria.
authors.forename: S
authors.initials: S
authors.lastname: Ecke
authors.forename: M
authors.initials: M
authors.lastname: Geiger
authors.forename: I
authors.initials: I
authors.lastname: Resch
authors.forename: I
authors.initials: I
authors.lastname: Jerabek
authors.forename: L
authors.initials: L
authors.lastname: Sting
authors.forename: M
authors.initials: M
authors.lastname: Maier
authors.forename: B R
authors.initials: BR
authors.lastname: Binder
mesh_terms: D000577:Amides / Q000378:metabolism; D001798:Blood Proteins / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D006868:Hydrolysis; D007610:Kallikreins / Q000037:antagonists & inhibitors* / Q000652:urine; D007700:Kinetics; D011486:Protein C / Q000378:metabolism*; D018046:Protein C Inhibitor; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D020840:Tissue Kallikreins
chemical_list: D000577:Amides; D001798:Blood Proteins; D011486:Protein C; D018046:Protein C Inhibitor; D015842:Serine Proteinase Inhibitors; D006493:Heparin; D007610:Kallikreins; D020840:Tissue Kallikreins