Open AccessWhy patients with THBD c.1611C>A (p.Cys537X) nonsense mutation have high levels of soluble thrombomodulin?
Author(s) -
Yohann Jourdy,
Nathalie Enjolras,
Sandra Le Quellec,
JeanClaude Bordet,
Claude Négrier,
Christine Vinciguerra,
Yesim Dargaud
Publication year - 2017
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0188213
Subject(s) - microbiology and biotechnology , mutant , transfection , thrombomodulin , western blot , mutation , proteases , mutagenesis , biology , chemistry , cell culture , thrombin , biochemistry , immunology , enzyme , gene , genetics , platelet
Background Recently our group has described a new autosomal dominant bleeding disorder characterized by very high plasma levels of soluble thrombomodulin (TM). The THBD c.1611C>A (p.Cys537X) mutation in heterozygous state was found in the propositus. This mutation leads to the synthesis of a truncated TM which has lost the last three amino-acids of the transmembrane domain and the cytoplasmic tail. Objective We investigated the mechanism responsible for TM shedding in endothelial cells with THBD c.1611C>A mutation. Methods Complementary DNA of TM wild type (TM-WT) was incorporated into a pcDNA3.1 vector for transient transfection in COS-1 cells. Mutagenesis was performed to create the c.1611C