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J. Biol. Chem., Vol. 280, Issue 15, 14462-14468, April 15, 2005

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Evidence for a Role of ADAM17 (TACE) in the Regulation of Platelet Glycoprotein V^*

Tamer Rabie, Amrei Strehl, Andreas Ludwig¶, and Bernhard Nieswandt||

From the Vascular Biology, Rudolf Virchow Center, Deutsche Forschungsgemeinschaft Research Center for Experimental Biomedicine, University of Würzburg, 97078 Würzburg, Germany, and ^¶Institute of Biochemistry, Christian Albrecht University Kiel, 24098 Kiel, Germany

Received for publication, January 3, 2005 , and in revised form, February 2, 2005.

	ABSTRACT

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

 
Glycoprotein V (GPV) is a subunit of the GPIb-IX-V receptor for von Willebrand factor and thrombin and has been shown to modulate platelet responses to the two strongest physiological agonists, thrombin and collagen. Thrombin directly cleaves GPV from the platelet surface, yielding a 69-kDa fragment GPV f1 of unknown function. We show here that a 82-kDa fragment of GPV is shed from the platelet surface upon cellular activation with phorbol 12-myristate 13-acetate or the collagen-related peptide. This shedding was inhibited by the broad range metalloproteinase inhibitor GM6001, the two potent ADAM17 inhibitors GW280264X and TAPI-2, and was absent in mice lacking functional ADAM17 (ADAM17 lacking Zn-binding domain; ADAM17^Zn/Zn). Furthermore, we show that recombinant ADAM17 ectodomain efficiently releases GPV from the platelet surface. GPV is known to be associated with the intracellular regulatory protein calmodulin, which has previously been shown to be involved in ADAM17-mediated shedding of L-selectin from the surface of leukocytes. As in these reports, inhibition of calmodulin led to rapid GPV shedding from the platelet surface, a process that was again blocked by GM6001 or ADAM17 inhibitors and that was absent in ADAM17^Zn/Zn mice. Inhibition of outside-in signaling through GPIIb/IIIa did not significantly affect GPV shedding, excluding an essential role of this pathway for the regulation of ADAM17 activity. These results demonstrate that GPV is cleaved upon agonist-induced platelet activation and show that ADAM17 is the major enzyme mediating this process.

	INTRODUCTION

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

 
Platelet adhesion and aggregation at sites of vascular injury is crucial to limit post-traumatic blood loss but may also harm tissue by occluding diseased vessels. The membrane glycoprotein (GP)¹ Ib-IX-V complex plays a central role in these events in that it mediates the initial platelet tethering to the damaged vessel wall under conditions of elevated shear by interacting with collagen-bound von Willebrand factor (1). The receptor complex consists of four distinct polypeptides, GPIb (M_r = 143,000), Ib (M_r = 22,000), V (M_r = 83,000), and IX (M_r = 20,000), in 1:1:0.5:1 stoichiometry with 25,000 copies per platelet. GPs Ib, V, and IX are structurally related and belong to the family of leucine-rich glycoproteins (2). The receptor complex is associated with the regulatory cytoplasmic protein calmodulin, which is involved in the free Ca²⁺ uptake upon platelet activation (3), but its exact role in GPIb-IX-V function is unclear.

The importance of the GPIb-IX-V complex is emphasized by the study of the Bernard-Soulier syndrome, an inherited bleeding disorder in which the complex is congenitally missing or dysfunctional because of mutations in the genes encoding GPIb or GPIX (2). Likewise, targeted deletion of the GPIb (4) or GPIb (5) genes in mice reproduces the Bernard-Soulier phenotype, as characterized by thrombocytopenia, giant platelets, and massively prolonged bleeding times. Although the essential role of GPIb-IX for normal platelet function is firmly established, the function of GPV in the receptor complex is only partly understood. GPV is merely loosely attached to GPIb-IX and, in contrast to GPs Ib and IX, is not essential for the expression of the receptor complex on the platelet surface. By contrast, GPIb and IX are required for expression of GP V (2). GPV-deficient mice have normal platelet size and counts, and the cells display an unaltered von Willebrand factor binding activity (6, 7).

Nevertheless, recent studies have revealed that GPV plays a role in different pathways of platelet activation. On the one hand, GPV was found to serve as a collagen receptor and to facilitate GPVI-dependent platelet-collagen interactions (8). On the other hand, GPV seems to act as a negative modulator of thrombin-induced platelet activation, in that cleavage of the protein unmasks GPIb-IX and allows binding of thrombin to GPIb (9). These observations suggest that regulation of GPV expression levels may play a role in determining the pathway of platelet activation in vivo, where several agonists act in concert during thrombus formation. However, knowledge about how the number of GPV molecules is regulated on the platelet surface is limited. It is well known that thrombin directly cleaves GPV, releasing a 69-kDa soluble fragment (GPV f1) of unknown function. In addition, elastase and calpain are capable of cleaving GPV as well as releasing 75-and 82-kDa fragments, respectively, but the significance of these observations is unresolved.

Proteolysis of membrane proteins is a well characterized regulatory process in a number of systems, including adhesion molecules, cytokines, growth factors, and their receptors. In most cases, shedding of ectodomains is mediated by membrane-anchored metalloproteinases (reviewed in Refs. 10–12). Calmodulin has been shown to play a role in shedding of L-selectin from leukocytes (13), a process that is mediated by tumor necrosis factor--converting enzyme (ADAM17), which belongs to the ADAM (a disintegrin and metalloproteinase) family of zinc-dependent proteinases. The importance of ADAM17-mediated ectodomain shedding is highlighted by the perinatal lethality of mice lacking the functional enzyme (14).

In the current study, we investigated the regulation of GPV in human and mouse platelets. We show that agonist-induced platelet activation or inhibition of calmodulin results in metalloproteinase-dependent shedding of an 82-kDa fragment of the receptor from the platelet surface. Furthermore, using mice lacking functional ADAM17 (ADAM17 lacking Zn-binding domain; ADAM17^Zn/Zn), we identify ADAM17 as the responsible sheddase.

	EXPERIMENTAL PROCEDURES

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

 
Mice—NMRI wild-type mice 6 to 10 weeks of age were obtained from Charles River Laboratories (Sulzfeld, Germany) and kept in our animal facilities. ADAM17^+/Zn mice (14) were kindly provided by Dr. Roy Black at Amgen (Seattle, WA) and crossed to obtain ADAM17^Zn/Zn mice.

Reagents—EZ-Link sulfo-NHS-LC-biotin (Pierce), Tetramethylbenzidine (Europa Bioproducts Ltd, Cambridge, UK), Sepharose G, (Amersham Biosciences), ADP, phorbol 12-myristate 13-acetate (PMA), high molecular weight heparin (Sigma), -thrombin (Roche Molecular Biochemicals), the broad range metalloproteinase inhibitor GM6001, the calmodulin inhibitors W7 and W13 (Calbiochem), human recombinant tumor necrosis factor--converting enzyme (ADAM17; R&D Systems, Minneapolis, MN), hirudin (Roche Diagnostics), streptavidin-HRP (DakoCytomation Denmark A/S, Glostrup, Denmark), ECL solution (Amersham Biosciences), and sulfuric acid (Applichem, Darmstadt, Germany) were purchased. Collagen-related peptide (CRP) was kindly provided by Steve Watson, and the specific metalloproteinase inhibitor GW280264X was kindly provided by GlaxoSmithKline (Stevenage, UK).

Antibodies—JON/A, which preferentially binds to activated murine GPIIb/IIIa was purchased from emfret Analytics (Würzburg, Germany). Anti-human GPV and anti-human GPIb were purchased from Immunotools (Friesoythe, Germany). All other antibodies were generated, produced, and modified in our laboratories: p0p6 (anti-murine GPIX), DOM1 and DOM2 (anti-murine GPV), and p0p1 (anti-human GPIb) (15, 16).

Platelet Preparation—Mice were bled under ether anesthesia from the retro-orbital plexus. Blood was collected in a tube containing 10% (v/v) 1 µM sodium citrate or 7.5 units/ml heparin, and platelet-rich plasma (prp) was obtained by centrifugation at 300 x g for 10 min at room temperature (RT). For washed platelets, prp was centrifuged at 1000 x g for 8 min at room temperature and the pellet was resupended twice in modified Tyrode-HEPES buffer (134 mM NaCl, 0.34 mM Na₂HPO₄, 2.9 mM KCL, 12 mM NaHCO₃, 20 mM HEPES, 5 mM glucose, and 0.35% bovine serum albumin, pH 6.6) in the presence of prostacyclin (0.1 µg/ml) and apyrase (0.02 units/ml). Platelets were then resuspended in the same buffer (pH 7.0; 0.02 units/ml apyrase) and incubated at 37 °C for at least 30 min before use. Isolated platelets did not show any signs of activation as shown by flow cytometry (staining for P-selectin and surface-expressed fibrinogen). Because ADAM17^Zn/Zn mice die perinatally (14), blood was collected from mutant and control mice immediately after birth.

Prepapartion of Washed Human Platelets—Platelet-rich plasma was prepared from TBS-EDTA anticoagulated blood obtained from aspirin free healthy volunteers and platelets were washed by sequential centrifugation as described previously (17). The cells were finally suspended in Tyrode's buffer and adjusted to 3 x 10⁵ platelets/ml.

Platelet Activation—Washed platelets were resuspended at a concentration of 0.5 x 10⁶ platelets/ml in modified Tyrode's buffer with 1 µM CaCl₂. The activators and inhibitors were added at the following concentrations: 2.5 µg/ml CRP, 5 µM ADP, 0.1 µg/ml PMA, 0.2 units/ml thrombin, 100 µM GM6001, 200 µM W13, 200 µM GW280264X, and incubated for 10 min (or 2 h for W13) at 37 °C. Platelets were then directly centrifuged at 400 x g for 5 min to obtain cell-free supernatants for ELISA and immunoprecipitation or analyzed by flow cytometry.

Platelet Treatment with Recombinant ADAM17—Platelets were washed twice in Tyrode's buffer in the presence of prostacyclin (0.1 µg/ml) and apyrase (0.02 units/ml) and finally resuspended in the absence of prostacyclin and apyrase to a concentration of 0.5 x 10⁹ platelets/ml. Recombinant ADAM17 (dissolved at a concentration of 100 µg/ml in 25 mM Tris, pH 9.0, containing 2.5 µM ZnCl₂ and 0.005% Brij according to the manufacturer's instructions) was added to the platelet suspension at the indicated concentrations and incubated for 1 h at 37 °C. Thereafter, the cells were analyzed by flow cytometry.

Flow Cytometry—Washed platelets (2 x 10⁶ in Tyrode's buffer and 1 mM CaCl₂) were incubated with fluorophore-conjugated monoclonal antibodies (10 µg/ml) for 10 min at RT, the reaction was stopped with 500 µl of PBS, and the samples were immediately analyzed on a FAC-SCalibur (BD Biosciences).

Aggregometry—To determine platelet aggregation, light transmission was measured using prp (200 µl with 0.5 x 10⁶ platelets/µl) and washed platelets. Transmission was recorded on a Fibrintimer 4 channel aggregometer (APACT Laborgeräte und Analysensysteme, Hamburg, Germany) over 10 min and was expressed as arbitrary units with 100% transmission adjusted with plasma. Platelet aggregation was induced by addition of PMA (0.1 µg/ml), CRP (2.5 µg/ml), U46619 [GenBank] (0.1 µM), and ADP (5 µM). Thrombin-induced aggregation was performed with washed platelets (200 µl with 0.5 x 10⁶ platelets/µl in Tyrode's buffer, 1 µmol/liter CaCl₂). For GIIb/IIIa inhibition, JON/A (50 µg/ml) was added 3 min before addition of the activators.

ELISA—To detect cleaved GPV molecule with the ELISA system, platelets were treated with the indicated activators and the supernatants (from 300 µl of 0.5 x 10⁹ platelets) were cleared by 10-min centrifugation at 15,000 x g. For GIIb/IIIa blocking, JON/A (50 µg/ml) was added 3 min before addition of the activators. Supernatants were transferred onto GPV-coated (DOM1; 20 µg/ml) ELISA plates, incubated for 1 h at 37 °C, and washed three times with washing buffer (1 x PBS and 0.1% Tween). Secondary antibody (DOM2-HRP) was added at a concentration of 2.5 µg/ml followed by 1 h incubation at 37 °C and detected with tetramethylbenzidine. The reaction was stopped with 0.5 M sulfuric acid and analyzed by WinRead.

Immunoprecipitation—Washed platelets were surface-labeled with EZ-Link sulfo-NHS-LC-biotin (25 µg/ml in PBS) and subsequently incubated with different reagents/agonists for the indicated times and then centrifuged (2000 x g; 10 min). Supernatants were then collected and incubated with 10 µg of DOM1 (anti-GPV), p0p4 (anti-GPIb), or EDL1 (anti-GPIIIa) and 25 µl of protein G-Sepharose (Amersham Biosciences) overnight at 4 °C. Samples were separated by 12% SDS-PAGE and transferred onto a polyvinylidene difluoride membrane. The membrane was incubated with streptavidin-HRP (1 µg/ml) for 1 h after blocking. After extensive washing, biotinylated proteins were visualized by ECL.

	RESULTS AND DISCUSSION

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

 
To study GPV regulation on the platelet surface, we stimulated human platelets with PMA (100 ng/ml), ADP (5 µM), or CRP (2,5 µg/ml) and determined surface levels of GPV by flow cytometry. CRP activates the cells through the GPVI/FcR-chain complex (18), the major platelet collagen receptor (19). As a positive control, platelets were incubated with thrombin (0.1 units/ml), which directly cleaves the receptor. GPV was down-regulated in response to PMA to a similar extent as in the thrombin control, whereas a weaker effect was observed in response to CRP. In contrast, ADP, which is only a weak platelet agonist, did not induce significant down-regulation of GPV (Fig. 1A).

View larger version (27K): [in this window] [in a new window]   FIG. 1. GPV down-regulation in human platelets is metalloproteinase-dependent. Washed human platelets were incubated for 15 min at 37 °C in the absence (shaded area) or presence (black line) of the indicated agonists, stained with FITC-labeled anti-GPV antibody (A) or anti-GPIb antibody (B) in the absence or presence of GM6001 (100 µM) (C) and analyzed immediately on a FACScalibur cell-sorting apparatus (BD Biosciences). B and C, data shown are the mean ± S.D. of six separate experiments and are presented as percentage of mean fluorescence determined at t = 0.

PMA- or CRP-induced shedding of GPV occurred independently of thrombin activity in that it was unaltered in the presence of the thrombin inhibitor hirudin (data not shown). In contrast, the broad spectrum metalloproteinase inhibitor GM6001 markedly blocked GPV shedding in response to PMA and CRP but not thrombin (Fig. 1C). Very similar results were obtained when GPV regulation was studied in murine platelets. In these cells, PMA induced virtually complete down-regulation of the receptor, whereas the effect of CRP was weaker and ADP did not alter GPV surface levels (Fig. 2A). GPIX surface levels remained virtually unchanged under all conditions, confirming that GPV shedding is not accompanied by down-regulation of the entire GPIbIX-V complex from the cell surface (Fig. 2A). As in human platelets, GM6001 almost completely inhibited GPV shedding in response to PMA or CRP but not thrombin (Fig. 2B). Immunoprecipitation experiments demonstrated that the metalloproteinase-generated fragment of GPV has a molecular mass of 82 kDa and thereby differs significantly from GPV f1 (69 kDa) released by thrombin (Fig. 2C). As a control, immunoprecipitations were also performed with an anti-GPIIIa antibody, which did not yield a band under any condition (Fig. 2C). These results demonstrated that platelet activation by thrombin or PMA and CRP induces limited proteolysis of GPV via different mechanisms. PMA- or CRP-stimulated cleavage involves a platelet-derived metalloproteinase and occurs near the transmembrane domain.

View larger version (35K): [in this window] [in a new window]   FIG. 2. Metalloproteinase- and thrombin-mediated GPV cleavage leads to the generation of different soluble GPV variants. Washed mouse platelets were incubated with the indicated agonists for 15 min at 37 °C, stained with FITC-labeled anti-GPV or anti-GPIX antibody (A) in the presence or absence of GM6001 (B), and analyzed immediately on a FACScalibur. The data shown are the mean ± S.D. of six separate experiments and are presented as percentage of mean fluorescence determined at t = 0. C, surface-biotinylated mouse platelets were incubated with the indicated agonists for 15 min at 37 °C in the absence or presence of GM6001 (100 µM). Thereafter, GPV and GPIIIa were immunoprecipitated from the supernatants of the cells. Immunoprecipitates were separated by SDS-PAGE under reducing conditions, blotted onto a polyvinylidene difluoride membrane, and visualized using streptavidin-HRP and ECL. The data shown are representative of three identical experiments.

Zn/Zn

View larger version (32K): [in this window] [in a new window]   FIG. 3. Inhibition of ADAM17 blocks GPV shedding from the platelet surface. A, whole platelet proteins were separated by SDS-PAGE under reducing conditions and immunoblotted with anti-ADAM17 antibodies. B, human (left) and mouse (right) platelets were activated in presence or absence of TAPI-2 (100 µM) or GW280264X (10 µM), stained with FITC-labeled anti-GPV antibody and analyzed immediately on a FACScalibur apparatus. Data shown are the mean ± S.D. of six separate experiments and are presented as percetnage of mean fluorescence determined at t = 0. C, washed mouse platelets were incubated with the indicated concentrations of recombinant human ADAM17 for 1 h at 37 °C, stained with FITC-labeled anti-GPV antibody and analyzed immediately on a FACScalibur apparatus. Where indicated, the experiment was performed in the presence of GM6001 (100 µM). Data shown are the mean ± S.D. of three separate experiments.

View larger version (27K): [in this window] [in a new window]   FIG. 4. GPV shedding is abolished in ADAM17Zn/Zn mice. Platelets from wild-type or ADAM17Zn/Zn mice were incubated in the absence (shaded area) or presence (black line) of the indicated agonists for 15 min at 37 °C, stained with FITC-conjugated anti-GPV antibody, and analyzed immediately on a FACScalibur. The results shown are representative of four independent experiments.

Zn/Zn

View larger version (27K): [in this window] [in a new window]   FIG. 5. Inhibition of calmodulin induces metalloproteinase-dependent shedding of GPV in mouse and human platelets. A, washed human and mouse platelets were incubated in the presence or absence of GM6001, with W13 (200 µM) for 2 h at 37 °C, stained with FITC-labeled anti-GPV antibody, and analyzed immediately on a FACScalibur apparatus. The data shown are the mean ± S.D. of six separate experiments. B, washed platelets from wild-type or ADAM17Zn/Zn mice were incubated with W13 (200 µM) for 2 h at 37 °C, stained with FITC-labeled anti-GPV antibody, and analyzed immediately on a FACScalibur apparatus. The data shown are the mean ± S.D. of four separate experiments. C, surface-biotinylated mouse platelets were incubated with W13 (200 µM) for 2 h at 37 °C in presence or absence of GM6001 (100 µM). Thereafter, GPV and GPIIIa were immunoprecipitated from the supernatants of the cells. Immunoprecipitates were separated by SDS-PAGE under reducing conditions, blotted onto a polyvinylidene difluoride membrane, and visualized using streptavidin-HRP and ECL. The data shown are representative of three identical experiments.

View larger version (16K): [in this window] [in a new window]   FIG. 6. GPV shedding occurs independently of GPIIbIIIa outside-in signaling. Washed mouse platelets (thrombin) or heparinized prp (PMA) were incubated with the indicated activators in absence or presence (*) of JON/A (50 µg/ml) and (A) light transmission was recorded on a Fibrintimer 4 channel aggregometer. B, GPV levels in the supernatants were determined by ELISA as described under "Experimental Procedures." Results shown are the mean ± S.D. of six individual experiments.

ADAM 17 is a well known sheddase that cleaves a variety of receptors, including L-selectin, VCAM-1 (14, 33, 34), and, as shown very recently, platelet GPIb (35). As in the regulation of these molecules, the release of GPV from the platelet surface can be stimulated by the phorbol ester PMA through activation of ADAM17, a pathway that seems to be conserved across multiple cell types. This is also supported by the observation that calmodulin plays a central role in the regulation of the shedding of GPV and L-selectin. Calmodulin is known to bind to the endodomain of both receptors, and prevention of such interaction by calmodulin inhibitors stimulates ectodomain shedding (Fig. 5) (13, 36). Very recent evidence suggests that similar mechanisms may be involved in the cellular regulation of the major platelet collagen receptor GPVI (26). Besides its interaction with receptors, calmodulin also acts as a cellular intermediate of multiple Ca²⁺ actions and acts to interfere with the protein kinase C pathway (37, 38). Although protein kinase C itself is a potent activator of ADAM17, protein kinase C-independent pathways of ADAM17 activation have been reported (36, 39). Although the pathways leading to ADAM17 activation may be conserved, the sequences cleaved by the enzyme in different proteins are highly variable, suggesting that it recognizes more complex motifs and/or sequences distal to the cleavage site. It remains to be elucidated how ADAM17 becomes activated and what exactly determines the substrate specificity of the enzyme (40).

In conclusion, we show that the endogenous metalloproteinase ADAM17 is the major sheddase for GPV on the platelet surface. The shedding process can be regulated via calmodulin inhibition and is independent of outside-in signaling through the GPIIb/IIIa complex. The physiological relevance of this shedding process needs to be further elucidated regarding both the impact of GPV shedding on the platelet itself and the biological properties of the GPV fragment released from the membrane.

	FOOTNOTES

 
^* This work was supported in part by the Deutsche Forschungsgemeinschaft Grants Ni556/4-1 (to B. N.) and LU 869/1-1 (to A. L.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this work.

^|| Heisenberg Fellow of the Deutsche Forschungsgemeinschaft. To whom correspondence should be addressed. Tel.: 49-931-201-48996; Fax: 49-931-201-48123; E-mail: bernhard.nieswandt{at}virchow.uniwuerzburg.de.

¹ The abbreviations used are: GP, glycoprotein; ADAM, a disintegrin and metalloproteinase; PMA, phorbol 12-myristate 13-acetate; HRP, horseradish peroxidase; CRP, collagen-related peptide; prp, plateletrich plasma; ELISA, enzyme-linked immunosorbent assay; FITC, fluorescein isothiocyanate; TAPI-2, TNF- protease inhibitor 2.

	ACKNOWLEDGMENTS

 
We thank S. Hartmann for excellent technical assistance.

	REFERENCES

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

 

1. Savage, B., Almus-Jacobs, F., and Ruggeri, Z. M. (1998) Cell 94, 657–666[CrossRef][Medline] [Order article via Infotrieve]
2. Andrews, R. K., Gardiner, E. E., Shen, Y., Whisstock, J. C., and Berndt, M. C. (2003) Int. J. Biochem. Cell Biol. 35, 1170–1174[CrossRef][Medline] [Order article via Infotrieve]
3. Andrews, R. K., Munday, A. D., Mitchell, C. A., and Berndt, M. C. (2001) Blood 98, 681–687[Abstract/Free Full Text]
4. Ware, J., Russell, S., and Ruggeri, Z. M. (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 2803–2808[Abstract/Free Full Text]
5. Kato, K., Martinez, C., Russell, S., Nurden, P., Nurden, A., Fiering, S., and Ware, J. (2004) Blood 104, 2339–2344[Abstract/Free Full Text]
6. Kahn, M. L., Diacovo, T. G., Bainton, D. F., Lanza, F., Trejo, J., and Coughlin, S. R. (1999) Blood 94, 4112–4121[Abstract/Free Full Text]
7. Ramakrishnan, V., Reeves, P. S., DeGuzman, F., Deshpande, U., Ministri-Madrid, K., DuBridge, R. B., and Phillips, D. R. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 13336–13341[Abstract/Free Full Text]
8. Moog, S., Mangin, P., Lenain, N., Strassel, C., Ravanat, C., Schuhler, S., Freund, M., Santer, M., Kahn, M., Nieswandt, B., Gachet, C., Cazenave, J. P., and Lanza, F. (2001) Blood 98, 1038–1046[Abstract/Free Full Text]
9. Ramakrishnan, V., DeGuzman, F., Bao, M., Hall, S. W., Leung, L. L., and Phillips, D. R. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 1823–1828[Abstract/Free Full Text]
10. Blobel, C. P. (2002) Inflamm. Res. 51, 83–84[CrossRef][Medline] [Order article via Infotrieve]
11. Moss, M. L., and Lambert, M. H. (2002) Essays Biochem. 38, 141–153[Medline] [Order article via Infotrieve]
12. Black, R. A., Doedens, J. R., Mahimkar, R., Johnson, R., Guo, L., Wallace, A., Virca, D., Eisenman, J., Slack, J., Castner, B., Sunnarborg, S. W., Lee, D. C., Cowling, R., Jin, G., Charrier, K., Peschon, J. J., and Paxton, R. (2003) Biochem. Soc. Symp. 39–52
13. Kahn, J., Walcheck, B., Migaki, G. I., Jutila, M. A., and Kishimoto, T. K. (1998) Cell 92, 809–818[CrossRef][Medline] [Order article via Infotrieve]
14. Peschon, J. J., Slack, J. L., Reddy, P., Stocking, K. L., Sunnarborg, S. W., Lee, D. C., Russell, W. E., Castner, B. J., Johnson, R. S., Fitzner, J. N., Boyce, R. W., Nelson, N., Kozlosky, C. J., Wolfson, M. F., Rauch, C. T., Cerretti, D. P., Paxton, R. J., March, C. J., and Black, R. A. (1998) Science 282, 1281–1284[Abstract/Free Full Text]
15. Bergmeier, W., Rackebrandt, K., Schroder, W., Zirngibl, H., and Nieswandt, B. (2000) Blood 95, 886–893[Abstract/Free Full Text]
16. Nieswandt, B., Bergmeier, W., Rackebrandt, K., Gessner, J. E., and Zirngibl, H. (2000) Blood 96, 2520–2527[Abstract/Free Full Text]
17. Cazenave, J. P., Ohlmann, P., Cassel, D., Eckly, A., Hechler, B., and Gachet, C. (2004) Methods Mol. Biol. 272, 13–28[Medline] [Order article via Infotrieve]
18. Kehrel, B., Wierwille, S., Clemetson, K. J., Anders, O., Steiner, M., Knight, C. G., Farndale, R. W., Okuma, M., and Barnes, M. J. (1998) Blood 91, 491–499[Abstract/Free Full Text]
19. Nieswandt, B., and Watson, S. P. (2003) Blood 102, 449–461[Abstract/Free Full Text]
20. Fox, J. E. (1994) Blood Coagul. Fibrinolysis 5, 291–304[Medline] [Order article via Infotrieve]
21. Hundhausen, C., Misztela, D., Berkhout, T. A., Broadway, N., Saftig, P., Reiss, K., Hartmann, D., Fahrenholz, F., Postina, R., Matthews, V., Kallen, K. J., Rose-John, S., and Ludwig, A. (2003) Blood 102, 1186–1195[Abstract/Free Full Text]
22. Abel, S., Hundhausen, C., Mentlein, R., Schulte, A., Berkhout, T. A., Broadway, N., Hartmann, D., Sedlacek, R., Dietrich, S., Muetze, B., Schuster, B., Kallen, K. J., Saftig, P., Rose-John, S., and Ludwig, A. (2004) J. Immunol. 172, 6362–6372[Abstract/Free Full Text]
23. Crowe, P. D., Walter, B. N., Mohler, K. M., Otten-Evans, C., Black, R. A., and Ware, C. F. (1995) J. Exp. Med. 181, 1205–1210[Abstract/Free Full Text]
24. Bennett, T. A., Edwards, B. S., Sklar, L. A., and Rogelj, S. (2000) J. Immunol. 164, 4120–4129[Abstract/Free Full Text]
25. Matala, E., Alexander, S. R., Kishimoto, T. K., and Walcheck, B. (2001) J. Immunol. 167, 1617–1623[Abstract/Free Full Text]
26. Gardiner, E. E., Arthur, J. F., Kahn, M. L., Berndt, M. C., and Andrews, R. K. (2004) Blood
27. Hidaka, H., and Tanaka, T. (1983) Methods Enzymol. 102, 185–194[Medline] [Order article via Infotrieve]
28. Tanaka, T., Ohmura, T., and Hidaka, H. (1983) Pharmacology 26, 249–257[CrossRef][Medline] [Order article via Infotrieve]
29. Andre, P., Prasad, K. S., Denis, C. V., He, M., Papalia, J. M., Hynes, R. O., Phillips, D. R., and Wagner, D. D. (2002) Nat. Med. 8, 247–252[CrossRef][Medline] [Order article via Infotrieve]
30. Prasad, K. S., Andre, P., He, M., Bao, M., Manganello, J., and Phillips, D. R. (2003) Proc. Natl. Acad. Sci. U. S. A. 100, 12367–12371[Abstract/Free Full Text]
31. Bergmeier, W., Schulte, V., Brockhoff, G., Bier, U., Zirngibl, H., and Nieswandt, B. (2002) Cytometry 48, 80–86[CrossRef][Medline] [Order article via Infotrieve]
32. Azorsa, D. O., Moog, S., Ravanat, C., Schuhler, S., Follea, G., Cazenave, J. P., and Lanza, F. (1999) Thromb. Haemost. 81, 131–138[Medline] [Order article via Infotrieve]
33. Walcheck, B., Alexander, S. R., St Hill, C. A., and Matala, E. (2003) J. Leukoc. Biol. 74, 389–394[Abstract/Free Full Text]
34. Garton, K. J., Gough, P. J., Philalay, J., Wille, P. T., Blobel, C. P., Whitehead, R. H., Dempsey, P. J., and Raines, E. W. (2003) J. Biol. Chem. 278, 37459–37464[Abstract/Free Full Text]
35. Bergmeier, W., Piffath, C. L., Cheng, G., Dole, V. S., Zhang, Y., von Andrian, U. H., and Wagner, D. D. (2004) Circ. Res. 95, 677–683[Abstract/Free Full Text]
36. Diaz-Rodriguez, E., Esparis-Ogando, A., Montero, J. C., Yuste, L., and Pandiella, A. (2000) Biochem. J. 346, 359–367
37. Massague, J., and Pandiella, A. (1993) Annu. Rev. Biochem. 62, 515–541[CrossRef][Medline] [Order article via Infotrieve]
38. Chakravarthy, B., Morley, P., and Whitfield, J. (1999) Trends Neurosci. 22, 12–16[CrossRef][Medline] [Order article via Infotrieve]
39. Pandiella, A., and Massague, J. (1991) J. Biol. Chem. 266, 5769–5773[Abstract/Free Full Text]
40. Black, R. A. (2002) Int. J. Biochem. Cell Biol. 34, 1–5[CrossRef][Medline] [Order article via Infotrieve]

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