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PNAS 103 (38): 13932-13937

Copyright © 2006 by the National Academy of Sciences.


Disulfide isomerization switches tissue factor from coagulation to cell signaling

Jasimuddin Ahamed*,{dagger}, Henri H. Versteeg*,{dagger}, Marjolein Kerver*, Vivien M. Chen{ddagger}, Barbara M. Mueller§, Philip J. Hogg{ddagger}, and Wolfram Ruf*

*Department of Immunology, The Scripps Research Institute, SP258, 10550 North Torrey Pines Road, La Jolla, CA 92037; {ddagger}Centre for Vascular Research, University of New South Wales, Sydney 2052, Australia; and §La Jolla Institute for Molecular Medicine, San Diego, CA 92121

Communicated by Earl W. Davie, University of Washington, Seattle, WA, July 27, 2006

Received for publication June 7, 2006.

Abstract: Cell-surface tissue factor (TF) binds the serine protease factor VIIa to activate coagulation or, alternatively, to trigger signaling through the G protein-coupled, protease-activated receptor 2 (PAR2) relevant to inflammation and angiogenesis. Here we demonstrate that TF·VIIa-mediated coagulation and cell signaling involve distinct cellular pools of TF. The surface-accessible, extracellular Cys186–Cys209 disulfide bond of TF is critical for coagulation, and protein disulfide isomerase (PDI) disables coagulation by targeting this disulfide. A TF mutant (TF C209A) with an unpaired Cys186 retains TF·VIIa signaling activity, and it has reduced affinity for VIIa, a characteristic of signaling TF on cells with constitutive TF expression. We further show that PDI suppresses TF coagulant activity in a nitric oxide-dependent pathway, linking the regulation of TF thrombogenicity to oxidative stress in the vasculature. Furthermore, a unique monoclonal antibody recognizes only the noncoagulant, cryptic conformation of TF. This antibody inhibits formation of the TF·PAR2 complex and TF·VIIa signaling, but it does not prevent coagulation activation. These experiments delineate an upstream regulatory mechanism that controls TF function, and they provide initial evidence that TF·VIIa signaling can be specifically inhibited with minimal effects on coagulation.

Key Words: allosteric disulfide • protein disulfide • isomerase • S-nitrosylation • G protein-coupled receptor

{dagger}J.A. and H.H.V. contributed equally to this work.

Author contributions: J.A., H.H.V., P.J.H., and W.R. designed research; J.A., H.H.V., M.K., V.M.C., and B.M.M. performed research; P.J.H. contributed new reagents/analytic tools; B.M.M., P.J.H., and W.R. analyzed data; and J.A., H.H.V., B.M.M., and W.R. wrote the paper.

Conflict of interest statement: No conflicts declared.

To whom correspondence should be addressed. E-mail: ruf{at}

© 2006 by The National Academy of Sciences of the USA

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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
Rapid activation of monocyte tissue factor by antithymocyte globulin is dependent on complement and protein disulfide isomerase.
F. Langer, B. Spath, C. Fischer, M. Stolz, F. A. Ayuk, N. Kroger, C. Bokemeyer, and W. Ruf (2013)
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
Thioredoxin and Thioredoxin Reductase Control Tissue Factor Activity by Thiol Redox-dependent Mechanism.
P. Wang, Y. Wu, X. Li, X. Ma, and L. Zhong (2013)
J. Biol. Chem. 288, 3346-3358
   Abstract »    Full Text »    PDF »
Protein-disulfide Isomerase Regulates the Thyroid Hormone Receptor-mediated Gene Expression via Redox Factor-1 through Thiol Reduction-Oxidation.
S. Hashimoto and S. Imaoka (2013)
J. Biol. Chem. 288, 1706-1716
   Abstract »    Full Text »    PDF »
New Fundamentals in Hemostasis.
H. H. Versteeg, J. W. M. Heemskerk, M. Levi, and P. H. Reitsma (2013)
Physiol Rev 93, 327-358
   Abstract »    Full Text »    PDF »
Murine tissue factor coagulant activity is critically dependent on the presence of an intact allosteric disulfide.
L. G. van den Hengel, S. Osanto, P. H. Reitsma, and H. H. Versteeg (2013)
Haematologica 98, 153-158
   Abstract »    Full Text »    PDF »
Protein Disulfide Isomerase Is Required for Platelet-derived Growth Factor-induced Vascular Smooth Muscle Cell Migration, Nox1 NADPH Oxidase Expression, and RhoGTPase Activation.
L. A. Pescatore, D. Bonatto, F. L. Forti, A. Sadok, H. Kovacic, and F. R. M. Laurindo (2012)
J. Biol. Chem. 287, 29290-29300
   Abstract »    Full Text »    PDF »
T granules in human platelets function in TLR9 organization and signaling.
J. N. Thon, C. G. Peters, K. R. Machlus, R. Aslam, J. Rowley, H. Macleod, M. T. Devine, T. A. Fuchs, A. S. Weyrich, J. W. Semple, et al. (2012)
J. Cell Biol. 198, 561-574
   Abstract »    Full Text »    PDF »
Tissue factor and glycoprotein C on herpes simplex virus type 1 are protease-activated receptor 2 cofactors that enhance infection.
M. R. Sutherland, W. Ruf, and E. L. G. Pryzdial (2012)
Blood 119, 3638-3645
   Abstract »    Full Text »    PDF »
The Michael Mason prize: Pathogenic antiphospholipid antibodies, stressed out antigens and the deployment of decoys.
Y. Ioannou (2012)
Rheumatology 51, 32-36
   Abstract »    Full Text »    PDF »
Lateral self-association of VWF involves the Cys2431-Cys2453 disulfide/dithiol in the C2 domain.
T. Ganderton, J. W. H. Wong, C. Schroeder, and P. J. Hogg (2011)
Blood 118, 5312-5318
   Abstract »    Full Text »    PDF »
Complete abolishment of coagulant activity in monomeric disulfide-deficient tissue factor.
L. G. van den Hengel, B. Kocaturk, P. H. Reitsma, W. Ruf, and H. H. Versteeg (2011)
Blood 118, 3446-3448
   Full Text »    PDF »
Global effects of fluvastatin on the prothrombotic status of patients with antiphospholipid syndrome.
C. Lopez-Pedrera, P. Ruiz-Limon, M. A. Aguirre, N. Barbarroja, C. Perez-Sanchez, P. Buendia, I.-C. Rodriguez-Garcia, A. Rodriguez-Ariza, E. Collantes-Estevez, F. Velasco, et al. (2011)
Ann Rheum Dis 70, 675-682
   Abstract »    Full Text »    PDF »
Cell-derived vesicles exposing coagulant tissue factor in saliva.
R. J. Berckmans, A. Sturk, L. M. van Tienen, M. C. L. Schaap, and R. Nieuwland (2011)
Blood 117, 3172-3180
   Abstract »    Full Text »    PDF »
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J. Disse, H. H. Petersen, K. S. Larsen, E. Persson, N. Esmon, C. T. Esmon, L. Teyton, L. C. Petersen, and W. Ruf (2011)
J. Biol. Chem. 286, 5756-5767
   Abstract »    Full Text »    PDF »
Endothelium-derived but not platelet-derived protein disulfide isomerase is required for thrombus formation in vivo.
R. Jasuja, B. Furie, and B. C. Furie (2010)
Blood 116, 4665-4674
   Abstract »    Full Text »    PDF »
Ero1{alpha} Is Expressed on Blood Platelets in Association with Protein-disulfide Isomerase and Contributes to Redox-controlled Remodeling of {alpha}IIb{beta}3.
M. Swiatkowska, G. Padula, L. Michalec, M. Stasiak, S. Skurzynski, and C. S. Cierniewski (2010)
J. Biol. Chem. 285, 29874-29883
   Abstract »    Full Text »    PDF »
Naturally occurring free thiols within {beta}2-glycoprotein I in vivo: nitrosylation, redox modification by endothelial cells, and regulation of oxidative stress-induced cell injury.
Y. Ioannou, J.-Y. Zhang, F. H. Passam, S. Rahgozar, J. C. Qi, B. Giannakopoulos, M. Qi, P. Yu, D. M. Yu, P. J. Hogg, et al. (2010)
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   Abstract »    Full Text »    PDF »
Extracellular protein disulfide isomerase regulates coagulation on endothelial cells through modulation of phosphatidylserine exposure.
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Blood 116, 993-1001
   Abstract »    Full Text »    PDF »
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H. Kothari, L. V. M. Rao, and U. R. Pendurthi (2010)
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   Full Text »    PDF »
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   Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »
Proteomics in antiphospholipid syndrome: a review.
M. Cuadrado, M. Aguirre, N. Barbarroja, M. Khamashta, and C. Lopez-Pedrera (2010)
Lupus 19, 385-388
   Abstract »    PDF »
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R. R. Bach and D. Monroe (2009)
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   Full Text »    PDF »
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F. Schaffner and W. Ruf (2009)
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   Abstract »    Full Text »    PDF »
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S. Butenas, T. Orfeo, and K. G. Mann (2009)
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   Abstract »    Full Text »    PDF »
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Blood 114, 4738-4740
   Abstract »    Full Text »    PDF »
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S. Carta, P. Castellani, L. Delfino, S. Tassi, R. Vene, and A. Rubartelli (2009)
J. Leukoc. Biol. 86, 549-555
   Abstract »    Full Text »    PDF »
In vitro and in vivo evidence for shear-induced activation of latent transforming growth factor-{beta}1.
J. Ahamed, N. Burg, K. Yoshinaga, C. A. Janczak, D. B. Rifkin, and B. S. Coller (2008)
Blood 112, 3650-3660
   Abstract »    Full Text »    PDF »
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H. H. Versteeg, F. Schaffner, M. Kerver, L. G. Ellies, P. Andrade-Gordon, B. M. Mueller, and W. Ruf (2008)
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   Abstract »    Full Text »    PDF »
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H. P. H. Liang and P. J. Hogg (2008)
Blood 112, 912-913
   Full Text »    PDF »
Response: Tissue factor de-encryption: the cell model system.
U. R. Pendurthi and L. V. M. Rao (2008)
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   Full Text »    PDF »
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Blood 111, 3295-3296
   Full Text »    PDF »
Factor Xa Binding to Annexin 2 Mediates Signal Transduction via Protease-Activated Receptor 1.
G. Bhattacharjee, J. Ahamed, R. Pawlinski, C. Liu, N. Mackman, W. Ruf, and T. S. Edgington (2008)
Circ. Res. 102, 457-464
   Abstract »    Full Text »    PDF »
The Haemostatic Role of Tissue Factor Pathway Inhibitor.
J. T.B. Crawley and D. A. Lane (2008)
Arterioscler Thromb Vasc Biol 28, 233-242
   Abstract »    Full Text »    PDF »
Inhibition of tissue factor signaling suppresses tumor growth.
H. H. Versteeg, F. Schaffner, M. Kerver, H. H. Petersen, J. Ahamed, B. Felding-Habermann, Y. Takada, B. M. Mueller, and W. Ruf (2008)
Blood 111, 190-199
   Abstract »    Full Text »    PDF »
Encryption remains cryptic.
J. H. Morrissey (2007)
Blood 110, 3822-3823
   Full Text »    PDF »
Tissue factor activation: is disulfide bond switching a regulatory mechanism?.
U. R. Pendurthi, S. Ghosh, S. K. Mandal, and L. V. M. Rao (2007)
Blood 110, 3900-3908
   Abstract »    Full Text »    PDF »
CCAAT/Enhancer-binding Protein-beta Participates in Insulin-responsive Expression of the Factor VII Gene.
A. A. Jackson, K. R. Cronin, R. Zachariah, and J. A. Carew (2007)
J. Biol. Chem. 282, 31156-31165
   Abstract »    Full Text »    PDF »
Tissue Factor Coagulant Function Is Enhanced by Protein-disulfide Isomerase Independent of Oxidoreductase Activity.
H. H. Versteeg and W. Ruf (2007)
J. Biol. Chem. 282, 25416-25424
   Abstract »    Full Text »    PDF »
Tissue factor trafficking in fibroblasts: involvement of protease-activated receptor-mediated cell signaling.
S. K. Mandal, U. R. Pendurthi, and L. V. M. Rao (2007)
Blood 110, 161-170
   Abstract »    Full Text »    PDF »
The Diversity of Bioactive Proteins in Australian Snake Venoms.
G. W. Birrell, S. T. H. Earl, T. P. Wallis, P. P. Masci, J. de Jersey, J. J. Gorman, and M. F. Lavin (2007)
Mol. Cell. Proteomics 6, 973-986
   Abstract »    Full Text »    PDF »
Stimulation of P2 (P2X7) receptors in human dendritic cells induces the release of tissue factor-bearing microparticles.
M. Baroni, C. Pizzirani, M. Pinotti, D. Ferrari, E. Adinolfi, S. Calzavarini, P. Caruso, F. Bernardi, and F. Di Virgilio (2007)
FASEB J 21, 1926-1933
   Abstract »    Full Text »    PDF »
Role of Protease Activated Receptor 1 and 2 Signaling in Hypoxia-Induced Angiogenesis.
H. Uusitalo-Jarvinen, T. Kurokawa, B. M. Mueller, P. Andrade-Gordon, M. Friedlander, and W. Ruf (2007)
Arterioscler Thromb Vasc Biol 27, 1456-1462
   Abstract »    Full Text »    PDF »
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S. Ghosh, U. R. Pendurthi, A. Steinoe, C. T. Esmon, and L. V. M. Rao (2007)
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   Abstract »    Full Text »    PDF »
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N. Wajih, S. M Hutson, and R. Wallin (2007)
J. Biol. Chem. 282, 2626-2635
   Abstract »    Full Text »    PDF »
Cryptic messages: Is noncoagulant tissue factor reserved for cell signaling?.
E. Camerer and J. Trejo (2006)
PNAS 103, 14259-14260
   Full Text »    PDF »

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