Back to Search
Start Over
Filamin-A is required for the incorporation of tissue factor into cell-derived microvesicles
- Source :
- Thrombosis and Haemostasis. 111:647-655
- Publication Year :
- 2014
- Publisher :
- Georg Thieme Verlag KG, 2014.
-
Abstract
- SummaryWe previously reported that the incorporation of tissue factor (TF) into cell-derived microvesicles (MVs) is regulated by the phosphorylation of the cytoplasmic domain of TF. Since the cytoskeletal protein filamin-A is known to bind to the cytoplasmic domain of TF in a phosphorylation-dependent manner, the involvement of filamin-A in the incorporation of TF into MVs was examined. Endothelial cells were transfected to express TF, whereas MDA-MB-231 cells were used to examine endogenously expressed TF. MV release was induced by activating protease-activated receptor-2 (PAR2). Partial suppression of filamin-A expression using two different filamin-A siRNA sequences resulted in significant reductions in the incorporation of TF antigen into MVs as determined by TF-ELISA and western blot analysis, and was reflected in reduced thrombin-generation and FXa-generation capacities of these MVs. Deletion of the cytoplasmic domain of TF also resulted in reduced incorporation of TF into MVs, whereas the suppression of filamin-A expression had no additional effect on the incorporation of truncated TF into MVs. Partial suppression of filamin-A expression had no effect on the number and size distribution of the released MVs. However, >90% suppression of filamin-A expression resulted in increased MV release, possibly as a result of increased instability of the plasma membrane and underlying cytoskeleton. In conclusion, the presence of filamin-A appears to be essential for the incorporation of TF into MVs following PAR2 activation, but is not required for the process of MV formation and release following PAR2 activation.
- Subjects :
- 0301 basic medicine
animal structures
Filamins
macromolecular substances
030204 cardiovascular system & hematology
Biology
Protein Engineering
Filamin
Thromboplastin
03 medical and health sciences
Tissue factor
0302 clinical medicine
Western blot
Cell-Derived Microparticles
medicine
Humans
Receptor, PAR-2
Particle Size
RNA, Small Interfering
Cytoskeleton
Cells, Cultured
Sequence Deletion
Secretory Pathway
medicine.diagnostic_test
Thrombin
Endothelial Cells
Hematology
Transfection
Molecular biology
Microvesicles
Protein Structure, Tertiary
Cell biology
030104 developmental biology
Cytoplasm
Factor Xa
Phosphorylation
Subjects
Details
- ISSN :
- 2567689X and 03406245
- Volume :
- 111
- Database :
- OpenAIRE
- Journal :
- Thrombosis and Haemostasis
- Accession number :
- edsair.doi.dedup.....8eacb0fc3db595a080cc1044a66f56f7