3.Laboratory assessment of von Willebrand factor for classification of von Willebrand disease.
Young Woo SON ; Kyung Chae KYE ; Hyun Chun SHIN ; Hong Bock LEE ; Do Yeun OH ; Seon Yang PARK ; Byeong Kook KIM ; Noe Kyeong KIM
Korean Journal of Hematology 1993;28(2):345-350
No abstract available.
Classification*
;
von Willebrand Diseases*
;
von Willebrand Factor*
4.Study on the impact of Trp1707Ser mutation on the binding mechanism of rF VIII light chain with VWF.
Kun CHI ; Yanyan SHAO ; Yeling LU ; Jing DAI ; Qiulan DING ; Xuefeng WANG ; Hongli WANG
Chinese Journal of Hematology 2014;35(11):995-999
OBJECTIVETo disclose the impact of Trp1707Ser mutation on the binding mechanism of rFVIII light chain (rFVIII LC) with VWF.
METHODSUsing long-chain PCR technique, we constructed rFVIII LC plasmids of both wild type and Trp1707Ser mutant type. BL21 competent cells were used for protein expression. Gradient renaturation was employed to refold protein. SDS-PAGE and Western blot were performed to identify the molecular weight of expressed protein. GST-Sefinose was used for protein purification and surface plasmon resonance (SPR) was employed to detect binding of B-domain-deleted rFVIII (BDD-rFVIII), wild and mutant rFVIII LC with VWF, respectively.
RESULTSThe results of SDS-PAGE and Western blot showed a molecular weight of 110×10(3) of expressed proteins, which were consistent with objective proteins. The expression quantity of wild type was higher than that of mutant type. A concentration-dependent combination of the 3 testing proteins with VWF was found. The KD value of BDDrFVIII (12.2) was lower than that of both rFVIII LCs (wild type 48.9 and mutant type 46.3), whereas there was no discrepancy between wild rFVIII LC and mutant rFVIII LC.
CONCLUSIONTrp1707Ser mutation didn't impact the binding of rFVIII LC expressed by BL21 competent cells with VWF. The heavy chain played a more important role in impacting the binding of FVIII with VWF.
Mutation ; von Willebrand Factor ; genetics
5.Research advance in von Willebrand factor.
Xue-Mei LI ; Miao JIANG ; Yi-Ming ZHAO
Journal of Experimental Hematology 2013;21(3):801-805
von Willebrand factor (vWF) is a multimeric glycoprotein exclusively synthesized in endothelial cells and megakaryocytes. It plays important roles in the primary and secondary haemostasis. Deficiency or dysfunction of vWF may cause von Willebrand disease (vWD), and overexpression of vWF may cause thrombosis. Making an intensive study on vWF will help us to understand the pathophysiology, diagnosis and treatment of vWF-related diseases, such as vWD, TTP, venous thrombosis, stroke, and so on. In this article, the regulation of vWF activity and its relation with diseases mentioned above are reviewed.
Humans
;
von Willebrand Diseases
;
blood
;
pathology
;
von Willebrand Factor
6.The Effect of VWF Propeptide on VWF Mutant in D1 Domain.
Xiu-Qun YU ; Zhen-Ni MA ; Jing LING ; Yun-Xiao ZHAO ; Jie YIN ; Zi-Qiang YU ; Chang-Geng RUAN
Journal of Experimental Hematology 2022;30(5):1541-1548
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.
HEK293 Cells
;
Humans
;
von Willebrand Diseases
;
von Willebrand Factor/metabolism*
10.Detection of Novel C4517G (Ser743Trp) Mutation in a Family with Type 2A von Willebrand Disease.
Kyung Soon SONG ; Hyun Kyung KIM ; Young Sook PARK
Korean Journal of Hematology 2003;38(4):274-278
Quantitative von Willebrand disease (VWD) are divided into partial deficiency (type 1) and total deficiency (type 3). Qualitative VWD are devided further into four subcategories (2A, 2B, 2M, 2N) based upon the major mechanism by which von Willebrand factor (VWF) function is impaired. Type 2A is characterized by the absence of large molecular weight VWF multimers and a number of mutations have been identified in the region encoding the A2 domain of VWF where a normal cleavage site is situated. Here, we report a case of type 2A VWD in a 5 year-old girl with a novel C4517G (Ser743Trp) mutation, which was also detected in her mother.
Child, Preschool
;
Female
;
Humans
;
Molecular Weight
;
Mothers
;
von Willebrand Disease, Type 2*
;
von Willebrand Diseases
;
von Willebrand Factor