The Effect of VWF Propeptide on VWF Mutant in D1 Domain.
10.19746/j.cnki.issn.1009-2137.2022.05.037
- Author:
Xiu-Qun YU
1
;
Zhen-Ni MA
1
;
Jing LING
2
;
Yun-Xiao ZHAO
1
;
Jie YIN
1
;
Zi-Qiang YU
1
;
Chang-Geng RUAN
3
Author Information
1. National Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Suzhou 215000, Jiangsu Province, China.
2. Children's Hospital of Soochow University, Department of Hematology and Oncology, Suzhou 215000, Jiangsu Province, China.
3. National Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Suzhou 215000, Jiangsu Province, China.E-mail: yuziqiang@suda.edu.cn.
- Publication Type:Journal Article
- Keywords:
biosynthesis;
secretion;
mutation;
von Willebrand factor;
von Willebrand factor propeptide
- MeSH:
HEK293 Cells;
Humans;
von Willebrand Diseases;
von Willebrand Factor/metabolism*
- From:
Journal of Experimental Hematology
2022;30(5):1541-1548
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate whether co-transfection of wild-type VWFpp with VWF mutant in D1 region is able to correct VWF defects in biosynthesis and secretion.
METHODS:Four VWF mutant plasmids were single transfected into HEK 293 cells, or co-transfected into HEK 293 cells with the wild type VWFpp plasmids. The VWF in supernatant and lysate of transfected cells were analyzed by ELISA, vertical VWF multimer electrophoresis. The retention of VWF in endoplasmic reticulum of transfected cells were detected by immunofluorescence confocal microscope.
RESULTS:In the vertical VWF multimer analysis, with co-expressing VWF mutant and VWFpp, the VWF multimer bands disappeared, and the VWF antigen in both supernatant and lysate of cells decreased, compared with the single expression of VWF mutant. Although the intracellular levels of VWF antigens decreased after co-expression, the retention rate of VWF mutant decreased in endoplasmic reticulum.
CONCLUSION:VWFpp can reduce the retention of VWF in endoplasmic reticulum, assists the transport of VWF between subcellular organelles. However, VWFpp inhibits the biosynthesis and secretion of VWF about the mutant in D1 domain.
