Expression of factor V on human umbilical vein endothelial cells is modulated by cell injury.

Annamalai, Anjanayaki E.; Stewart, Gwendolyn J.; Hansel, B C; Memoli, Matthew J.; Chiu, Hui Chong; Manuel, D W; Doshi, K; Colman, Robert W.

doi:10.1161/01.atv.6.2.196

Cited by 20 publications

(6 citation statements)

References 33 publications

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“…Cellular proteases with similar function exist in the endo-thelia! cell, which is also known to synthesize (51) and express FV (52,53). Rodgers et al (54) have demonstrated a 4-fold activation of FV in the absence of thrombin by a calcium dependent protease contained in vascular endothelial cells.…”

Section: Discussionmentioning

confidence: 99%

Thrombin-stimulated Human Platelets Express Molecular Forms of α-Granule Factor V (FV), which Differ from FVa

1994

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SummaryBinding of 125I-Fab fragments of chain-specific antibodies indicate that both heavy and light chains of a-granule factor Va (FVa) were externalized on the platelet membrane after stimulation with thrombin. Using a Mab against the activation peptide of factor V (FV), the epitope appears on the stimulated platelet surface. Half as much light chain and heavy chain (FVa) was expressed compared to the activation peptide, suggesting that expression of α-granule FV occurs after thrombin stimulation. Using an ELISA, we find that 32% of a-granule FV was released and 68% is retained in the platelet pellet. Immunoblots of platelets indicate that FV exists in 200 kDa und 150 kDa forms, representing incomplete cleavage, while the releasate demonstrates a more complete cleavage by proteases. We conclude that expression of α-granule FV is quantitatively greater than that released and exists in molecular forms which cannot be completely explained by the binding of FVa.

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Section: Discussionmentioning

confidence: 99%

Thrombin-stimulated Human Platelets Express Molecular Forms of α-Granule Factor V (FV), which Differ from FVa

1994

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“…Although endogenous synthesis of factor V by bovine aortic endothehium was reported by Cerveny et al (13) and Maruyama et al (14) found that a monoclonal antibody (mAb)2 to factor V/Va bound specifically to 15,000 sites on human umbilical vein endothelial cells (HUVEC), Annamalai et al (15) failed to confirm this observation on unperturbed endothelium and Jenny et al (16) did not detect any factor V message in these cells. Similarly, the proposed synthesis of factor V/Va by mononuclear cells (11) was challenged by the absence of factor V message in monocytic cells (16) and by studies of Kappelmayer et al (17), who failed to detect genuine factor V transcripts in monocytes using reverse-transcribed polymerase chain reaction combined with in situ hybridization strategy.…”

Section: Structure-function Of Factor Xamentioning

confidence: 98%

Xa receptor EPR‐1

Altieri

1995

FASEB j.

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Cellular inflammatory responses and early mechanisms of vascular injury are invariably associated with activation of blood coagulation and deposition of insoluble fibrin. This process occurs on vascular cell surfaces through the ability of the coagulation protease factor Xa to generate thrombin. However, experimental evidence accumulated during the past decade underscores how prothrombin activation is only one of the biological consequences of factor Xa assembly on vascular cells. Instead, binding of factor Xa to leukocytes, endothelium, and smooth muscle cells triggers complex pathways of intracellular signal transduction that participate, directly or indirectly, in the regulation of cellular growth. One of the cellular binding sites for factor Xa, designated effector cell protease receptor-1 (EPR-1), has recently emerged as a novel potential regulator of factor Xa-mediated mitogenic signaling. For its activation-dependent phenotype on leukocyte subsets, its ability to costimulate lymphocyte proliferation through release of intracellular second messengers, and its regulated cellular expression by alternative mRNA splicing, EPR-1 may influence vascular cell growth and aberrantly contribute to the earliest pathogenetic processes of vascular diseases.

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“…Consistent with the expression and function of EPR-1 on endothelium, 1 a synthetic peptide duplicating the interacting motif S 123 PGKPGNQNSKNEPP 137 dose-dependently inhibited thrombin formation on these cells in the absence of factor V/Va. That prothrombin activation on HUVEC could not be solely mediated by released factor V/Va (24) was suggested earlier by the absence of factor V/Va on normal unperturbed endothelium (25) and/or by the lack of factor V transcript in these cells (26). Moreover, cellular receptors for factor Xa, distinct from factor V/Va, were identified on HUVEC (27,28) and implicated in ligand processing (29) and intracellular signal transduction with release of endothelial cell mitogens (30).…”

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confidence: 89%

Molecular Dissection of Effector Cell Protease Receptor-1 Recognition of Factor Xa

Ambrosini

Altieri

1996

Journal of Biological Chemistry

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Assembly of coagulation and anticoagulation pathways occurs on vascular cells through the regulated ligand recognition of membrane protease receptors (1, 2). Belonging to structurally different and evolutionarily unrelated gene superfamilies, cell-surface receptors for thrombin (3), urokinase (4), protein S (5), or factor Xa (6) provide a controlled microenvironment for limited proteolytic activation of the clotting and fibrinolytic cascades (1, 2). Recent studies have also underscored the participation of protease receptors in pleiotropic mechanisms of vascular cell signal transduction, including transcription of activation-dependent genes (7), generation of intracellular second messengers (3), modulation of immune inflammatory responses (8), and cell proliferation (5, 9). Aberrations of protease-dependent signaling pathways may play a primary pathogenetic role in the establishment and progression of the atherothrombotic disease (9, 10) as well as in neoplastic transformation and tumor cell dissemination (5, 11).Binding of factor Xa to leukocytes (12) or endothelial cells 1 is contributed by effector cell protease receptor-1 (EPR-1), 2 a 337-amino acid single membrane-spanning glycoprotein lacking significant homologies to other protease receptors and expressed in a cell-type specific fashion by alternative mRNA splicing (13). Occupancy of EPR-1 with factor Xa participates in factor V/Va-independent mechanisms of prothrombin activation (6), generates intermediate product(s) of factor IX activation (14), or contributes an accessory pathway of lymphocyte costimulation (8).To gain insights into the potential pathophysiological relevance of EPR-1-factor Xa interaction in hemostasis and vascular cell signal transduction, we sought to identify the structural requirements implicated in receptor-ligand recognition. Using a eukaryotic expression strategy of EPR-1 chimeric constructs combined with site-directed mutagenesis and synthetic peptidyl mimicry, we have identified two spatially distinct regions in the EPR-1 extracellular domain separately involved in lymphocyte activation mechanisms or factor Xa binding and prothrombin activation. 2 The abbreviations used are: EPR-1, effector cell protease receptor-1; ICAM-1, intercellular adhesion molecule-1; PCR, polymerase chain reaction; bp, base pairs; CHO, Chinese hamster ovary; mAb monoclonal antibody; HUVEC, human umbilical vein endothelial cell(s). EXPERIMENTAL PROCEDURES Construction of EPR-1-ICAM-1 Chimeras-Three

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Expression of factor V on human umbilical vein endothelial cells is modulated by cell injury.

Cited by 20 publications

References 33 publications

Thrombin-stimulated Human Platelets Express Molecular Forms of α-Granule Factor V (FV), which Differ from FVa

Thrombin-stimulated Human Platelets Express Molecular Forms of α-Granule Factor V (FV), which Differ from FVa

Xa receptor EPR‐1

Molecular Dissection of Effector Cell Protease Receptor-1 Recognition of Factor Xa

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