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

Z. Bagoly, Ferenc Fazakas, I. Komáromi, G. Haramura, Eszter Tóth, L. Muszbek

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16 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)668-674
Number of pages7
JournalThrombosis and Haemostasis
Volume99
Issue number4
DOIs
Publication statusPublished - Apr 2008

Fingerprint

Factor XIII
Leukocyte Elastase
Thrombin
Transglutaminases
Valine
Factor XIIIa
Batroxobin
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Fibrin
Proteolysis
Arginine
Catalytic Domain
Fatty Acids
Western Blotting

Keywords

  • Elastase
  • Factor XIII
  • Factor XIII-A V34L polymorphism
  • Transglutaminase

ASJC Scopus subject areas

  • Hematology

Cite this

@article{6af68086937f4c6ea73c82046f33377c,
title = "Cleavage of factor XIII by human neutrophil elastase results in a novel active truncated form of factor XIII A subunit",
abstract = "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 = "Elastase, Factor XIII, Factor XIII-A V34L polymorphism, Transglutaminase",
author = "Z. Bagoly and Ferenc Fazakas and I. Kom{\'a}romi and G. Haramura and Eszter T{\'o}th and L. Muszbek",
year = "2008",
month = "4",
doi = "10.1160/TH07-09-0577",
language = "English",
volume = "99",
pages = "668--674",
journal = "Thrombosis and Haemostasis",
issn = "0340-6245",
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TY - JOUR

T1 - Cleavage of factor XIII by human neutrophil elastase results in a novel active truncated form of factor XIII A subunit

AU - Bagoly, Z.

AU - Fazakas, Ferenc

AU - Komáromi, I.

AU - Haramura, G.

AU - Tóth, Eszter

AU - Muszbek, L.

PY - 2008/4

Y1 - 2008/4

N2 - 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.

AB - 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.

KW - Elastase

KW - Factor XIII

KW - Factor XIII-A V34L polymorphism

KW - Transglutaminase

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U2 - 10.1160/TH07-09-0577

DO - 10.1160/TH07-09-0577

M3 - Article

C2 - 18392324

AN - SCOPUS:43749101177

VL - 99

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EP - 674

JO - Thrombosis and Haemostasis

JF - Thrombosis and Haemostasis

SN - 0340-6245

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