A novel congenital dysprothrombinemia leading to defective prothrombin maturation

Valeria Bafunno, Loredana Bury, Giovanni Luca Tiscia, Tiziana Fierro, Giovanni Favuzzi, Rocco Caliandro, Francesco Sessa, Elvira Grandone, Maurizio Margaglione, Paolo Gresele

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Abstract

Introduction: Prothrombin deficiency is a very rare disorder caused by mutations in the F2 gene that generate hypoprothrombinemia or dysprothrombinemia and is characterized by bleeding manifestations that can vary from clinically irrelevant to life-threatening. Aim: Here we characterize a patient with a novel missense mutation in F2, c.1090 T/A (p. Val322Glu), that causes severe dysprothrombinemia. Methods: Coagulation assays, prothrombin Western Blotting, FII activation by Ecarin, fibrinogen degradation products quantification and thrombin generation assay were carried out to assess prothrombin expression and function. PCR followed by direct sequencing was carried out to characterize the mutation. In silico analysis for missense variant and molecular modeling were applied to predict the mechanism that leads to dysprothrombinemia. Results and conclusions: The homozygous patient had a markedly prolonged prothrombin time, strongly reduced FII activity (0.82%) but normal antigen levels. In the thrombin generation assay the lag time and the peak height were unmeasurable, suggesting that the Val322Glu mutation results in the inability of the mutant prothrombin to be fully activated to thrombin. In fact, prothrombin activation by ecarin was defective, with a massive accumulation of the meizothrombin intermediate. Molecular modeling and dynamic simulation studies showed that the Val322Glu mutation interferes with protein flexibility at Arg271 and Arg320. This impairs the switch of the protein from zymogen to proteinase, thus preventing the formation of thrombin. Accumulated meizothrombin, however, maintains some fibrinogen-degrading activity, as shown by the formation of FDPs, and this probably explains the patient's mild bleeding phenotype.

Original languageEnglish
Pages (from-to)1135-1141
Number of pages7
JournalThrombosis Research
Volume134
Issue number5
DOIs
Publication statusPublished - 2014

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Keywords

  • Dysprothrombinemia
  • Factor II
  • Meizothrombin
  • Molecular modeling
  • Prothrombin activation

ASJC Scopus subject areas

  • Hematology
  • Medicine(all)

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