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Thromb Haemost 2017; 117(08): 1498-1508
DOI: 10.1160/TH17-02-0097
Coagulation and Fibrinolysis
Schattauer GmbH

Plasma carboxypeptidase U (CPU, CPB2, TAFIa) generation during in vitro clot lysis and its interplay between coagulation and fibrinolysis

Dorien Leenaerts
1   Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Wilrijk, Belgium
,
Jef Aernouts
2   Laboratory of Biomedical Physics, Department of Physics, University of Antwerp, Wilrijk, Belgium
,
Pieter Van Der Veken
3   Laboratory of Medicinal Chemistry, Department of Pharmaceutical Sciences, University of Antwerp, Wilrijk, Belgium
,
Yani Sim
1   Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Wilrijk, Belgium
,
Anne-Marie Lambeir
1   Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Wilrijk, Belgium
,
Dirk Hendriks
1   Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Wilrijk, Belgium
› Author Affiliations
Further Information

Publication History

Received: 13 February 2017

Accepted after major revision: 02 May 2017

Publication Date:
22 November 2017 (online)

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Summary

Carboxypeptidase U (CPU, CPB2, TAFIa) is a basic carboxypeptidase that is able to attenuate fibrinolysis. The inactive precursor procarboxypeptidase U is converted to its active form by thrombin, the thrombin-thrombomodulin complex or plasmin. The aim of this study was to investigate and characterise the time course of CPU generation in healthy individuals. In plasma of 29 healthy volunteers, CPU generation was monitored during in vitro clot lysis. CPU activity was measured by means of an enzymatic assay that uses the specific substrate Bz-o-cyano-Phe-Arg. An algorithm was written to plot the CPU generation curve and calculate the parameters that define it. In all individuals, CPU generation was biphasic. Marked inter-individual differences were present and a reference range was determined. The endogenous CPU generation potential is the composite effect of multiple factors. With respect to the first CPU activity peak characteristics, we found correlations with baseline proCPU concentration, proCPU Thr325Ile polymorphism, time to clot initiation and the clot lysis time. The second CPU peak related with baseline proCPU levels and with the maximum turbidity of the clot lysis profile. In conclusion, our method offers a technique to determine the endogenous CPU generation potential of an individual. The parameters obtained by the method quantitatively describe the different mechanisms that influence CPU generation during the complex interplay between coagulation and fibrinolysis, which are in line with the threshold hypothesis.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Thrombin activatable fibrinolysis inhibitor (proCPU, proCPB2, TAFI) - carboxypeptidases - carboxypeptidase B2 - clot lysis time - fibrinolysis

 

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