Extracellular protein disulfide isomerase regulates coagulation on endothelial cells through modulation of phosphatidylserine exposure.
Tissue factor (TF) is the cellular receptor for plasma protease factor VIIa (FVIIa), and the TF-FVIIa complex initiates coagulation in both hemostasis and thrombosis. Cell surface-exposed TF is mainly cryptic and requires activation to fully exhibit the procoagulant potential. Recently, the protein disulfide isomerase (PDI) has been hypothesized to regulate TF decryption through the redox switch of an exposed disulfide in TF extracellular domain. In this study, we analyzed PDI contribution to coagulation using an in vitro endothelial cell model. In this model, extracellular PDI is detected by imaging and flow cytometry. Inhibition of cell surface PDI induces a marked increase in TF procoagulant function, whereas exogenous addition of PDI inhibits TF decryption. The coagulant effects of PDI inhibition were sensitive to annexin V treatment, suggesting exposure of phosphatidylserine (PS), which was confirmed by prothrombinase assays and direct labeling. In contrast, exogenous PDI addition enhanced PS internalization. Analysis of fluorescent PS revealed that PDI affects both the apparent flippase and floppase activities on endothelial cells. In conclusion, we identified a new mechanism for PDI contribution to coagulation on endothelial cells, namely, the regulation of PS exposure, where PDI acts as a negative regulator of coagulation.
Popescu
Narcis I
NI
Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK 73104, USA.
Lupu
Cristina
C
Lupu
Florea
F
D017209:Apoptosis / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics; D001777:Blood Coagulation / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000201:enzymology; D042783:Endothelial Cells / Q000166:cytology / Q000201:enzymology*; D004789:Enzyme Activation / Q000502:physiology; D005110:Extracellular Space / Q000201:enzymology; D005434:Flow Cytometry; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D010718:Phosphatidylserines / Q000378:metabolism* / Q000494:pharmacology; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000378:metabolism*; D013925:Thromboplastin / Q000378:metabolism
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D010718:Phosphatidylserines; C054562:yellow fluorescent protein, Bacteria; D013925:Thromboplastin; C526504:PDIA2 protein, human; D019704:Protein Disulfide-Isomerases; D002118:Calcium
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