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Blood, 15 April 2004, Vol. 103, No. 8, pp. 3038-3044.
Prepublished online as a Blood First Edition Paper on December 30, 2003; DOI 10.1182/blood-2003-07-2399.
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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Lipid rafts are necessary for tonic inhibition of cellular tissue factor procoagulant activity
Dennis J. Dietzen,
Keith L. Page, and
Tina A. Tetzloff
From the Department of Pediatrics, Washington University, St Louis, MO.
A fraction of total cellular tissue factor procoagulant activity remains masked or "encrypted" in intact cells. Decryption of this activity partly involves the extracellular exposure of anionic phospholipids such as phosphatidylserine. Because of the potential association of tissue factor and phospholipid scramblase activity with lipid rafts, we have explored the role of lipid rafts in regulating factor VIIa/tissue factor activity. In HEK293 cells, tissue factor antigen was not stably associated with lipid rafts, yet disruption of rafts with methyl- -cyclodextrin resulted in a 3-fold stimulation of tissue factor procoagulant activity. Treatment with methyl--cyclodextrin was not associated with cytotoxicity and did not result in the exposure of additional tissue factor antigen. Factor VIIa/tissue factor activity decrypted with methyl--cyclodextrin was quantitatively similar to that obtained by using lytic concentrations of octyl glucoside but more sensitive to inhibition by cell surface tissue factor pathway inhibitor and the phospholipid binding protein, annexin V. Partial decryption of tissue factor was achieved with methyl--cyclodextrin prior to complete disruption of lipid rafts, suggesting the role of an enzyme localized to lipid rafts in the transbilayer transport of phosphatidylserine. We conclude that lipid rafts are required for the maintenance of cellular tissue factor in an encrypted state. (Blood. 2004;103:3038-3044)
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