Extracellular Signal-regulated Kinase (ERK) Activation Is Required for GP Ibα-dependent Endothelial Cell Migration

2001 ◽  
Vol 86 (12) ◽  
pp. 1555-1562 ◽  
Author(s):  
Jie Lian ◽  
Cezary Marcinkiewicz ◽  
Stefan Niewiarowski ◽  
Dorothy Beacham
Keyword(s):  
Endothelial Cell ◽  
Type I Collagen ◽  
Endothelial Cell Migration ◽  
Type I ◽  
Erk Activation ◽  
Extracellular Signal Regulated Kinase ◽  
Extracellular Signal ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Venom Proteins

SummaryThe GP Ib complex can participate in endothelial cell (EC) migration on von Willebrand factor (vWF) or the mixed matrix of vWF and type I collagen (vWF/collagen). In this study, viper venom proteins alboaggregin (albo) A or B blocked GP Ibα, and echistatin inhibited αvβ3 binding. Albo A, B and echistatin inhibited EC migration on vWF and vWF/collagen. Albo B or the anti-GP Ibα monoclonal antibody (mAb) 1b1 did not affect the migration of smooth muscle cells or fibroblasts, which lack GP Ib. EC also migrate on albo A- or albo B-coated dishes. PD98059, which blocks ERK activation, abolished EC migration on vWF, vWF/collagen, collagen or albo B. Soluble albo A or 1b1 dramatically inhibited ERK activation during EC migration on vWF or albo B. Echistatin inhibited ERK activation on vWF and vitronectin (VN), but not albo B. Thus, in addition to αvβ3, EC GP Ibα initiates ERK activation, and regulates ERK-induced EC migration on vWF.

scholarly journals Crotoxin Inhibits Endothelial Cell Functions in Two- and Three-dimensional Tumor Microenvironment

2021 ◽  
Vol 12 ◽  
Author(s):  
Ellen Emi Kato ◽  
Luciana Araújo Pimenta ◽  
Maíra Estanislau Soares de Almeida ◽  
Vanessa Olzon Zambelli ◽  
Marinilce Fagundes dos Santos ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Endothelial Cell ◽  
Actin Polymerization ◽  
Type I Collagen ◽  
Three Dimensional ◽  
Endothelial Cell Migration ◽  
Endothelial Cell Proliferation ◽  
Type I ◽  
Cell Functions ◽  
Extracellular Matrix Components

Antitumor property of Crotoxin (CTX), the major toxin from Crotalus durissus terrificus snake venom, has been demonstrated in experimental animal models and clinical trials. However, the direct action of this toxin on the significant events involved in neovascularization, which are essential for tumor growth and survival, has not been confirmed. This study investigated the effects of CTX on the key parameters of neovascularization in two- and three-dimensional culture models. Murine endothelial cell lines derived from thymus hemangioma (t.End.1) were treated at different concentrations of CTX (6.25–200 nM). Endothelial cell proliferation, cell adhesion, and actin cytoskeletal dynamics on laminin (10 µg/ml), type I collagen (10 µg/ml), and fibronectin (3 µg/ml) were evaluated along with the endothelial cell migration and formation of capillary-like tubes in 3D Matrigel. CTX concentration of 50 nM inhibited tube formation on 3D Matrigel and impaired cell adhesion, proliferation, and migration under both culture medium and tumor-conditioned medium. These actions were not accountable for the loss of cell viability. Inhibition of cell adhesion to different extracellular matrix components was related to the reduction of αv and α2 integrin distribution and cytoskeletal actin polymerization (F-actin), accompanied by inhibition of focal adhesion kinase (FAK), Rac1 (GTPase) signaling proteins, and actin-related protein 2/3 (Arp 2/3) complex. This study proved that CTX inhibits the major events involved in angiogenesis, particularly against tumor stimuli, highlighting the importance of the anti-angiogenic action of CTX in inhibition of tumor progression.

Integrin alpha 2 I-domain is a binding site for collagens

1995 ◽  
Vol 108 (4) ◽  
pp. 1629-1637 ◽  
Author(s):  
D. Tuckwell ◽  
D.A. Calderwood ◽  
L.J. Green ◽  
M.J. Humphries
Keyword(s):  
Type I Collagen ◽  
Type I ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Collagen Binding ◽  
Functional Antibodies ◽  
A Domains ◽  
Domain I ◽  
Von Willebrand ◽  
Willebrand Factor

Integrins alpha 1 beta 1 and alpha 2 beta 1 are major cellular receptors for collagens. The alpha 1 and alpha 2 subunits contain a approximately 200 amino acid inserted domain (I-domain) in their N-terminal region and, because of the homology between the I-domains and the collagen-binding A-domains of von Willebrand factor, it has been suggested that the I-domains might mediate the collagen-binding functions of alpha 1 beta 1 and alpha 2 beta 1. In order to fully investigate this hypothesis, we have generated recombinant human alpha 2 I-domain (r alpha 2I) by reverse transcriptase-polymerase chain reaction/bacterial expression and tested its ability to mediate the collagen-binding functions of alpha 2 beta 1. R alpha 2 I binds specifically to type I collagen in a concentration-dependent manner: binding is cation dependent and, like the complete receptor, is supported by magnesium and manganese ions but not by calcium ions. R alpha 2I is recognised by anti-functional anti-alpha 2 monoclonal antibodies 6F1, 5E8 and P1E6 in capture ELISAs, and anti-functional antibodies inhibited r alpha 2I-collagen binding. In addition, r alpha 2I inhibits cell spreading on collagen. R alpha 2I is therefore a collagen-binding domain and can account for many of the collagen-binding functions of integrin alpha 2 beta 1. We have also determined the collagen specificity of r alpha 2I and found that it binds types I, II and XI collagen.

scholarly journals Impact of fibronectin assembly on platelet thrombus formation in response to type I collagen and von Willebrand factor

Blood ◽  
2006 ◽  
Vol 108 (7) ◽  
pp. 2229-2236 ◽  
Author(s):  
Jaehyung Cho ◽  
Deane F. Mosher
Keyword(s):  
Shear Rate ◽  
Von Willebrand Factor ◽  
Type I Collagen ◽  
Thrombus Formation ◽  
Type I ◽  
Plasma Fibronectin ◽  
Platelet Thrombus ◽  
Von Willebrand ◽  
Fibronectin Assembly ◽  
Willebrand Factor

Abstract Plasma fibronectin enhances platelet thrombus formation on surfaces coated with collagen. We investigated the role of fibronectin assembly in this process. Platelets adherent to fibrillar type I collagen, but not platelets adherent to von Willebrand factor (VWF), supported assembly of plasma fibronectin under static conditions. At a shear rate of 1250 s–1, platelets adherent to collagen assembled coperfused plasma fibronectin and formed larger thrombi in a fibronectin-concentration–dependent manner, with a maximum effect at 250 μg/mL. Enhanced thrombus formation on collagen was blocked by a peptide that binds to the N-terminal region of fibronectin and inhibits fibronectin assembly. Cross-linking of fibronectin to collagen prior to exposure to platelets had no effect on thrombus formation. Collagen-induced platelet thrombus formation at a shear rate of 5000 s–1 required coperfusion with VWF and did not result in assembly of coperfused fibronectin. VWF-mediated increase in platelet thrombi on collagen was not enhanced and indeed was somewhat attenuated by coperfused fibronectin at a shear rate of 5000 s–1. These results indicate that, at moderately high but not very high shear rates, fibronectin assembly in platelet aggregates that form in response to collagen enhances thrombus formation and serves as an alternative to VWF-mediated enhancement.

scholarly journals EGFL6 Promotes Endothelial Cell Migration and Angiogenesis through the Activation of Extracellular Signal-regulated Kinase

2011 ◽  
Vol 286 (25) ◽  
pp. 22035-22046 ◽  
Author(s):  
Shek Man Chim ◽  
An Qin ◽  
Jennifer Tickner ◽  
Nathan Pavlos ◽  
Tamara Davey ◽  
...  
Keyword(s):  
Cell Migration ◽  
Endothelial Cell ◽  
Endothelial Cell Migration ◽  
Extracellular Signal Regulated Kinase ◽  
Extracellular Signal

Characterization of W1745C and S1783A: 2 novel mutations causing defective collagen binding in the A3 domain of von Willebrand factor

Blood ◽  
2009 ◽  
Vol 114 (16) ◽  
pp. 3489-3496 ◽  
Author(s):  
Anne F. Riddell ◽  
Keith Gomez ◽  
Carolyn M. Millar ◽  
Gillian Mellars ◽  
Saher Gill ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Type I Collagen ◽  
Binding Activity ◽  
Von Willebrand Disease ◽  
Site Directed Mutagenesis ◽  
Type Iii Collagen ◽  
Type I ◽  
Collagen Binding ◽  
Von Willebrand ◽  
Willebrand Factor

AbstractInvestigation of 3 families with bleeding symptoms demonstrated a defect in the collagen-binding activity of von Willebrand factor (VWF) in association with a normal VWF multimeric pattern. Genetic analysis showed affected persons to be heterozygous for mutations in the A3 domain of VWF: S1731T, W1745C, and S1783A. One person showed compound heterozygosity for W1745C and R760H. W1745C and S1783A have not been reported previously. The mutations were reproduced by site-directed mutagenesis and mutant VWF expressed in HEK293T cells. Collagen-binding activity measured by immunosorbent assay varied according to collagen type: W1745C and S1783A were associated with a pronounced binding defect to both type I and type III collagen, whereas the principal abnormality in S1731T patients was a reduction in binding to type I collagen only. The multimer pattern and distribution of mutant proteins were indistinguishable from wild-type recombinant VWF, confirming that the defect in collagen binding resulted from the loss of affinity at the binding site and not impairment of high-molecular-weight multimer formation. Our findings demonstrate that mutations causing an abnormality in the binding of VWF to collagen may contribute to clinically significant bleeding symptoms. We propose that isolated collagen-binding defects are classified as a distinct subtype of von Willebrand disease.

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