Versican G3 domain promotes blood coagulation through suppressing the activity of tissue factor pathway inhibitor-1.
We have detected versican, a member of the large chondroitin sulfate proteoglycans, and its degraded C-terminal G3 fragments in human plasma and observed that the versican G3 domain promoted blood coagulation. Silencing G3 expression with small interfering RNA reduced the effect of G3 on coagulation. Plasma coagulation assays suggest that G3 enhances coagulation irrespective of its actions on platelets and white blood cells. To examine how versican affected blood coagulation, we used normal human plasma and different types of coagulation factor-deficient plasmas. The experiments indicated that versican enhanced coagulation through the extrinsic pathway, and that Factor VII was the target molecule. FVII activity assays showed that G3 activated FVII in the presence of plasma but not with purified FVII directly. Yeast two-hybrid, immunoprecipitation, and gel co-migration assays showed that G3 interacted with the tissue factor pathway inhibitor-1 (TFPI-1). TFPI-1 activity assays suggested that G3 inhibited TFPI-1 activity, allowing FVIIa and FXa to facilitate the coagulation process. G3-induced blood coagulation was further confirmed with a mouse model in a real-time manner. Taken together, these results indicate that versican may represent a new target for the development of therapies against atherosclerosis.
Zheng
Peng-Sheng
PS
Sunnybrook & Women's College Health Sciences Centre, University of Toronto, Ontario, Canada.
Reis
Marciano
M
Sparling
Cathy
C
Lee
Daniel Y
DY
La Pierre
David P
DP
Wong
Chung-Kwun Amy
CK
Deng
Zhaoqun
Z
Kahai
Shireen
S
Wen
Jianping
J
Yang
Burton B
BB
D020816:Amino Acid Motifs; D000818:Animals; D050197:Atherosclerosis; D001777:Blood Coagulation; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D011508:Chondroitin Sulfate Proteoglycans / Q000737:chemistry*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005822:Genetic Vectors; D006023:Glycoproteins / Q000737:chemistry; D006801:Humans; D047468:Immunoprecipitation; D037181:Lectins, C-Type / Q000737:chemistry*; D008074:Lipoproteins / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013997:Time Factors; D014162:Transfection; D020798:Two-Hybrid System Techniques; D053675:Versicans
D011508:Chondroitin Sulfate Proteoglycans; D017077:Culture Media, Conditioned; D006023:Glycoproteins; D037181:Lectins, C-Type; D008074:Lipoproteins; C508322:VCAN protein, human; C508323:Vcan protein, mouse; C051928:lipoprotein-associated coagulation inhibitor; D053675:Versicans