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Thromb Haemost 2008; 99(04): 668-674
DOI: 10.1160/TH07-09-0577
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Cleavage of factor XIII by human neutrophil elastase results in a novel active truncated form of factor XIII A subunit

Zsuzsa Bagoly
1   Clinical Research Center
2   Thrombosis, Haemostasis and Vascular Biology Research Group of the Hungarian Academy of Sciences, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary
,
Ferenc Fazakas
2   Thrombosis, Haemostasis and Vascular Biology Research Group of the Hungarian Academy of Sciences, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary
,
István Komáromi
2   Thrombosis, Haemostasis and Vascular Biology Research Group of the Hungarian Academy of Sciences, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary
,
Gizella Haramura
1   Clinical Research Center
,
Eszter Tóth
2   Thrombosis, Haemostasis and Vascular Biology Research Group of the Hungarian Academy of Sciences, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary
,
László Muszbek
1   Clinical Research Center
2   Thrombosis, Haemostasis and Vascular Biology Research Group of the Hungarian Academy of Sciences, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary
› Author AffiliationsFinancial support: This work was supported by grants from the Hungarian National Research Fund (OTKA-NKTH NI 69238), from the Hungarian Academy of Sciences (MTA 2006TKI227), from the Hungarian Ministry of Health and Social Affairs (ETT 406/2006) and from the National Office of Research and Technology (NKTH RET-06/2004).
Further Information

Publication History

Received: 24 September 2007

Accepted after major revision: 19 February 2008

Publication Date:
25 November 2017 (online)

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Summary

The first step in the activation of plasma factor XIII (FXIII) is the cleavage of R37-G38 bond in FXIIIA subunit (FXIII-A) by thrombin, which makes the subsequent formation of an active transglutaminase possible. No active truncated form of FXIII-A, other than G38-FXIII-A, has been identified. In contrast to thrombin, which has a preference toward arginine residues, human neutrophil elastase (HNE) cleaves peptide bonds at small side-chain aliphatic amino acids, preferably at valine. As there are several valine residues close to the thrombin cleavage-site, we tested if an active truncated FXIII-A was formed during fragmentation of FXIII by HNE. It was demonstrated by Western blotting and transglutaminase assay that HNE induced a limited cleavage of FXIII-A resulting in the activation of both plasma and cellular FXIII; the maximal transglutaminase activities were 52.5% and 67.4% of thrombin-activated FXIII, respectively. After the relatively rapid activation a much slower inactivation occurred. HNE-activated FXIII cross-linked fibrin γ- and α-chains in the clot formed by batroxobin moojeni. MALDI-TOF analysis of the cleaved fragments and N-terminal Edman degradation of the truncated protein identified V39-N40 as the primary cleavagesite and N40-FXIII-A as the active form. No primary cleavage occurred at V34, V35, V47, V50 residues. FXIII-A V34L polymorphism, which increases the rate of FXIII-A cleavage by thrombin, was without effect on FXIII activation by HNE. Molecular modeling located the primary HNE cleavage-site in the middle of the flexible and accessible Q32-L45 loop and showed that other neighboring valine residues were in less favorable position.

Keywords

Factor XIII - transglutaminase - elastase - factor XIII-A V34L polymorphism
 

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