The disulfide bond Cys2724-Cys2774 in the C-terminal cystine knot domain of von Willebrand factor is critical for its dimerization and secretion

Thrombosis Journal

Xiaoying Wang¹  Jingyu Zhang¹  Yuxin Zhang²  Yi Wu²  Aizhen Yang³  Fengwu Chen³  Yue Han³  Depei Wu³ 

[1] Department of Hematology, Key Laboratory of Hematology of Hebei Province, The Second Hospital of Hebei Medical University, 050000, Shijiazhuang, China;Department of Hematology, Key Laboratory of Hematology of Hebei Province, The Second Hospital of Hebei Medical University, 050000, Shijiazhuang, China;National Clinical Research Center for Hematologic Diseases, the First Affiliated Hospital, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Cyrus Tang Medical Institute, Soochow University, 215123, Suzhou, China;National Clinical Research Center for Hematologic Diseases, the First Affiliated Hospital, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Cyrus Tang Medical Institute, Soochow University, 215123, Suzhou, China;

关键词: von Willebrand factor (VWF);    von Willebrand disease (VWD);    cystine knot domain;    multimerization;    disulfide bond;   

DOI  :  10.1186/s12959-021-00348-w

来源: Springer

【 摘 要 】

BackgroundType 3 von Willebrand disease (VWD) exhibits severe hemorrhagic tendency with complicated pathogenesis. The C-terminal cystine knot (CTCK) domain plays an important role in the dimerization and secretion of von Willebrand factor (VWF). The CTCK domain has four intrachain disulfide bonds including Cys2724-Cys2774, Cys2739-Cys2788, Cys2750-Cys2804 and Cys2754-Cys2806, and the single cysteine mutation in Cys2739-Cys2788, Cys2750-Cys2804 and Cys2754-Cys2806 result in type 3 VWD, demonstrating the crucial role of these three disulfide bonds in VWF biosynthesis, however, the role of the remaining disulfide bond Cys2724-Cys2774 remains unclear.Method and resultsIn this study, by the next-generation sequencing we found a missense mutation a c.8171G>A (C2724Y) in the CTCK domain of VWF allele in a patient family with type 3 VWD. In vitro, VWF C2724Y protein was expressed normally in HEK-293T cells but did not form a dimer or secrete into cell culture medium, suggesting that C2724 is critical for the VWF dimerization, and thus for VWF multimerization and secretion.ConclusionsOur findings provide the first genetic evidence for the important role of Cys2724-Cys2774 in VWF biosynthesis and secretion. Therefore, all of the four intrachain disulfide bonds in CTCK monomer contribute to VWF dimerization and secretion.

【 授权许可】

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