Protein C Inhibitor from Human Plasma: Characterization of Native and Cleaved Inhibitor and Demonstration of Inhibitor Complexes with Plasma Kallikrein

Martin Laurell; Johan Stenflo

doi:10.1055/s-0038-1651022

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1989; 62(03): 885-891
DOI: 10.1055/s-0038-1651022

Original Article

Schattauer GmbH Stuttgart

Protein C Inhibitor from Human Plasma: Characterization of Native and Cleaved Inhibitor and Demonstration of Inhibitor Complexes with Plasma Kallikrein

Martin Laurell

The Department of Clinical Chemistry, Malmö General Hospital Malmö, Sweden

,

Johan Stenflo

The Department of Clinical Chemistry, Malmö General Hospital Malmö, Sweden

› Author Affiliations

Abstract

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Summary

Protein C inhibitor (PCI) was purified from human plasma using immunoaffinity chromatography and heparin Sepharose chromatography, a method that allowed the purification of active and inactive inhibitor. PCI purified from outdated plasma was inactive and either in complex with plasma kallikrein or proteo-lytically degraded. Sequence analysis of cleaved PCI and of complexes between PCI and activated protein C or urokinase identified the previously recognized inhibitor cleavage site Arg₃₅₄-Ser₃₅₅. Two additional cleavage sites were observed in the modified inhibitor i.e. Arg₃₅₇-Leu₃₅₈ and Arg₃₆₂-Leu₃₆₃ which probably represent secondary cleavage of the inhibitor. Furthermore the sequence analysis of the inhibitor, whether purified from fresh or outdated plasma, revealed that it was microheterogeneous in the NH₂-terminus as a result of cleavage by a trypsin like enzyme(s).

Keywords

Protein C - Protein C inhibitor - Plasma kallikrein - Urokinase - Heparin

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References

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