Inhibition of coagulation factors by recombinant barley serpin BSZx
- 30 September 1996
- journal article
- Published by Wiley in FEBS Letters
- Vol. 394 (2) , 165-168
- https://doi.org/10.1016/0014-5793(96)00940-4
Abstract
Barley serpin BSZx is a potent inhibitor of trypsin and chymotrypsin at overlapping reactive sites (Dahl, S.W., Rasmussen, S.K. and Hejgaard, J. (1996) J. Biol. Chem., in press). We have now investigated the interactions of BSZx with a range of serine proteinases from human plasma, pancreas and leukocytes, a fungal trypsin and three subtilisins. Thrombin, plasma kallikrein, factor VIIa/tissue factor and factor Xa were inhibited by BSZx at heparin independent association rates (k ass) of 4.5 × 103−1.3 × 105 M−1 s−1 at 22°C. Only factor Xa tunred a significant fraction of BSZx over as substrate. Complexes of these proteinase with BSZx resisted boiling in SDS, and amino acid sequencing showed that cleavage in the reactive center loop only occurred after P1 Arg. Activated protein C and leukocyte elastase were slowly inhibited by BSZx (k ass = 1−2 × 102 M−1 s−1) whereas factor XIIa, urokinase and tissue type plasminogen activator, plasmin and pancreas kallikrein and elastase were not or only weakly affected. The inhibition pattern with mammalian proteinases reveal a specificity of BSZx similar to that of antithrombin III. Trypsin from Fusarium was not inhibited while interaction with subtilisin Carlsberg and Novo was rapid but most BSZx was cleaved as a substrate. Identification of a monoclonal antibody specific for native BSZx indicate that complex formation and loop cleavage result in similar conformational changes.Keywords
This publication has 23 references indexed in Scilit:
- Interaction of subtilisins with serpinsProtein Science, 1996
- Characterization of common neoantigenic epitopes generated in plasminogen activator inhibitor-1 after cleavage of the reactive center loop or after complex formation with various serine proteinasesFEBS Letters, 1995
- Structural aspects of serpin inhibitionFEBS Letters, 1994
- Serpins from wheat grainFEBS Letters, 1994
- Evolutionary relationships among the serpinsPhilosophical Transactions Of The Royal Society B-Biological Sciences, 1993
- Chymotrypsin inhibitory activity of normal C1‐inhibitor and a P1 Arg to His mutant: Evidence for the presence of overlapping reactive centersProtein Science, 1993
- Alpha-2-Antiplasmin: a Serpin with Two Separate But Overlapping Reactive SitesScience, 1988
- A common neoepitope is created when the reactive center of C1-inhibitor is cleaved by plasma kallikrein, activated factor XII fragment, C1 esterase, or neutrophil elastase.Journal of Clinical Investigation, 1988
- Purification and properties of protein Z – a major albumin of barley endospermPhysiologia Plantarum, 1982
- p-Nitrophenyl-p′-guanidinobenzoate HCl: A new active site titrant for trypsinBiochemical and Biophysical Research Communications, 1967