Congenital afibrinogenemia: Intracellular retention of fibrinogen due to a novel W437G mutation in the fibrinogen Bβ-chain gene

Silvia Spena, Rosanna Asselta, Stefano Duga, Massimo Malcovati, Flora Peyvandi, Pier Mannuccio Mannucci, Maria Luisa Tenchini

Research output: Contribution to journalArticle

Abstract

Congenital afibrinogenemia is a rare autosomal recessive coagulation disorder characterised by hemorrhagic manifestations of variable entity and by severe plasma fibrinogen deficiency. Among the 31 afibrinogenemia-causing mutations so far reported, only 2 are missense mutations and both are located in the fibrinogen Bβ-chain gene. Direct sequencing of the fibrinogen gene cluster in two afibrinogenemic Iranian siblings revealed a novel homozygous T>G transversion in exon 8 (nucleotide position 8025) of the fibrinogen Bβ-chain gene. The resulting W437G missense mutation involves a highly conserved amino acid residue, located in the C-terminal globular D domain. The role of the W437G amino acid substitution on fibrinogen synthesis, folding, and secretion was assessed by in vitro expression experiments in COS-1 cells, followed by qualitative and quantitative analyses of intracellular and secreted mutant fibrinogen. Results of both pulse-chase experiments and enzyme-linked immunosorbent assays demonstrated intracellular retention of the mutant W437G fibrinogen and marked reduction of its secretion. These data, besides elucidating the pathogenetic role of the W437G mutation in afibrinogenemia, underline the importance of the Bβ-chain D domain in fibrinogen folding and secretion.

Original languageEnglish
Pages (from-to)87-94
Number of pages8
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Volume1639
Issue number2
DOIs
Publication statusPublished - Oct 15 2003

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Keywords

  • Congenital afibrinogenemia
  • Fibrinogen Bβ-chain
  • Missense mutation
  • Protein in vitro expression

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Biophysics

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