Thromb Haemost 2013; 110(01): 53-61
DOI: 10.1160/TH13-03-0184
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

The missense Thr211Pro mutation in the factor X activation peptide of a bleeding patient causes molecular defect in the clotting cascade

Qiulan Ding*
1   Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China
,
Yiping Shen*
2   Shanghai Children’s Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai, China
3   Department of Laboratory Medicine and Pathology, Children’s Hospital, Boston, Harvard Medical School, Massachusetts, USA
,
Likui Yang
4   Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, Saint Louis, Missouri, USA
,
Xuefeng Wang
1   Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China
,
Alireza R. Rezaie
4   Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, Saint Louis, Missouri, USA
› Author Affiliations
Further Information

Publication History

Received: 04 March 2013

Accepted after major revision: 21 April 2013

Publication Date:
30 November 2017 (online)

Summary

Factor X (FX) is a vitamin K-dependent coagulation zymogen, which upon activation to factor Xa assembles into the prothrombinase complex to activate prothrombin to thrombin. FX can be activated by either factor VIIa-tissue factor or factor IXa-factor VIIIa in extrinsic and intrinsic pathways, respectively. In this study, we identified a bleeding patient with moderate FX deficiency who exhibits a clotting defect only in the intrinsic pathway. Exome sequencing revealed that the patient carries a novel homozygous missense mutation that results in substitution of Thr211 with Pro in the activation peptide of FX. Thr211 is the site of an O-linked glycosylation in the activation peptide of FX. We postulated that the lack of this post-translational modification specifically impacts the activation of FX by intrinsic Xase, thereby impairing thrombin generation in the subject. To test this hypothesis, we expressed both wild-type FX and FX containing this mutation in mammalian cells and following the purification of the zymogens to homogeneity characterized their properties in both purified and plasma-based assay systems. Analysis of the results suggests that Thr211 to Pro substitution renders the FX mutant a poor substrate for both physiological activators, however, at physiological concentration of the substrate, the clotting defect manifest itself only in the intrinsic pathway, thus explaining the bleeding phenotype for the patient carrying this mutation.

* Q. Ding and Y. Shen contributed equally to this work.


 
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