The Disulfide Bond between Cys22 and Cys27 in the Protease Domain Modulate Clotting Activity of Coagulation Factor X

Fang Li; Changming Chen; Si-Ying Qu; Ming-Zhu Zhao; Xiaoling Xie; Xi Wu; Lei Li; Xuefeng Wang; Qiulan Ding; Qin Xu; Dong-Qing Wei; Wenman Wu

doi:10.1055/s-0039-1683442

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2019; 119(06): 871-881
DOI: 10.1055/s-0039-1683442

Coagulation and Fibrinolysis

Georg Thieme Verlag KG Stuttgart · New York

The Disulfide Bond between Cys22 and Cys27 in the Protease Domain Modulate Clotting Activity of Coagulation Factor X

Fang Li^*

¹State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China

,

Changming Chen^*

²Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

,

Si-Ying Qu

¹State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China

,

Ming-Zhu Zhao

³Instrumental Analysis Center, Shanghai Jiao Tong University, Shanghai, China

,

Xiaoling Xie

²Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

,

Xi Wu

²Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

,

Lei Li

²Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

,

Xuefeng Wang

²Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

⁴Faculty of Laboratory Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai, China

,

Qiulan Ding

²Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

,

Qin Xu

¹State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China

,

Dong-Qing Wei

¹State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China

,

Wenman Wu

²Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

⁴Faculty of Laboratory Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai, China

⁵Collabortive Innovation Center of Hematology, Shanghai Jiao Tong University School of Medicine, Shanghai, China

› Institutsangaben

Abstract

The Cys22-Cys27 disulfide bond of factor X (FX) protease domain is not conserved among coagulation factors and its contribution to the physiological haemostasis and implication in the pathogenesis of haemostatic and thrombotic disorders remain to be elucidated. Mutation p.Cys27Ser was identified in a pedigree of congenital FX deficiency and fluorescence labelling study of transiently transfected HEK293 cells showed accumulation of FX p.Cys27Ser within cell, indicating incompetent secretion partially responsible for the FX deficiency. The clotting activity of FX p.Cys27Ser was decreased to about 90% of wild-type, while amidolytic and pro-thrombinase activities (kcat/Km) determined with recombinant FXa mutant were 1.33- and 4.77-fold lower. Molecular dynamic simulations revealed no major change in global structure between FXa p.Cys27Ser and wild-type FXa; however, without the Cys22-Cys27 disulfide bond, the insertion of newly formed N terminal of catalytic domain after the activation cleavage is hindered, perturbing the conformation transition from zymogen to enzyme. The crystal structure of FXa shows that this disulfide bond is solvent accessible, indicating that its stability might be subject to the oxidation/reduction balance. As demonstrated with FX p.Cys27Ser here, Cys22-Cys27 disulfide bond may modulate FX clotting activity, with reduced FX pertaining less pro-coagulant activity.

Keywords

coagulation factor X - Cys27 - disulfide bond - oxidation/reduction balance - molecular dynamic simulations

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Referenzen

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