Handrková, Helena; Schroeder, Verena; Kohler, H P (2015). The activation peptide of coagulation factor XIII is vital for its expression and stability. Journal of thrombosis and haemostasis, 13(8), pp. 1449-1458. Wiley-Blackwell 10.1111/jth.13035
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BACKGROUND
The human activation peptide of factor XIII (AP-FXIII) comprises the first 37 amino acids of the N-terminus and holds the FXIII in an inactive state. FXIII is activated either proteolytically by cleavage of AP-FXIII by thrombin, or non-proteolytically by high calcium concentrations.
OBJECTIVE
To investigate the role of AP-FXIII in the expression and stability of FXIII.
METHODS
We cloned 13 FXIII variants with progressive truncations of AP-FXIII from the N-terminus (delN-FXIII-A), expressed them in mammalian cells, and measured their thermostability, activation, and transglutaminase activity. We also used in silico calculations to analyze the stability of hypothetical delN-FXIII dimers and to identify crucial motifs within AP-FXIII.
RESULTS
Variants with deletions longer than the first 10 amino acids and an R11Q point mutant were not expressed as proteins. In silico calculations indicated that the sequence (8) FGGR(12) R plays a substantial role in intersubunit interactions in FXIII-A2 homodimers. In agreement with this prediction, the temperature stability of delN-FXIII variants decreased with increasing length of deletion. These results may suggest a role of the N-terminus of AP-FXIII in dimer stability. Substantial sequence homology was found among activation peptides of vertebrate and even invertebrate (crustacean) FXIII-A orthologs, which further supports our conclusion.
CONCLUSIONS
We conclude that deletion of 11 or more N-terminal amino acids disrupts intersubunit interactions, which may prevent FXIII-A2 homodimer formation. Therefore, AP-FXIII plays an important role in the stability of the FXIII-A2 dimer.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Pavillon 52 > Forschungsgruppe Experimentelle Hämostase 04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Clinic of Haematology and Central Haematological Laboratory |
UniBE Contributor: |
Röss, Helena, Schröder, Verena |
Subjects: |
600 Technology > 610 Medicine & health |
ISSN: |
1538-7836 |
Publisher: |
Wiley-Blackwell |
Language: |
English |
Submitter: |
Verena Zwahlen |
Date Deposited: |
02 May 2016 11:49 |
Last Modified: |
02 Mar 2023 23:27 |
Publisher DOI: |
10.1111/jth.13035 |
PubMed ID: |
26083359 |
Uncontrolled Keywords: |
dimerization, factor XIII, factor XIII activation peptide, plasma transglutaminase, protein stability |
BORIS DOI: |
10.7892/boris.81596 |
URI: |
https://boris.unibe.ch/id/eprint/81596 |
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