Characterization of the bleeding disorder. (A) Pedigree of the affected family. The proband is indicated by an arrow. Family members with a history of trauma- and/or surgery-related bleeding are marked by closed symbols. (B-D) Thrombin generation experiments. Plasma was activated with the indicated concentrations of TF, 4 μM phospholipids, and 16.6 mM CaCl2. Thrombin generation was measured using calibrated automated thrombography in the patient’s plasma (black lines) or control plasma (gray lines). (D) Mixtures of the patient’s and control plasma in different proportions were activated with 1 pM TF, and the ETP was plotted as a function of the percentage of patient’s plasma in the mixtures. (E) Analysis of the whole exome sequencing data. The 64 885 variants identified in the patient’s DNA were filtered for minor allele frequency (MAF) <0.01 (based on the assumption that the mutation responsible for this bleeding disorder would be rare), for heterozygosity (because of the dominant inheritance pattern of the disorder), and for location in coagulation-related genes. These criteria yielded 8 candidate variants (table), which were further analyzed individually. (F) Schematic representation of the TM molecule anchored to the endothelial membrane. The position of the Cys537 residue is indicated.
