The binding of heparin to the extracellular matrix of endothelial cells up-regulates the synthesis of an antithrombotic heparan sulfate proteoglycan

The binding of heparin to the extracellular matrix of endothelial cells up-regulates the synthesis of an antithrombotic heparan sulfate proteoglycan

Autor Trindade, Edvaldo S. Google Scholar
Oliver, Constance Google Scholar
Jamur, Maria C. Google Scholar
Rocha, Hugo A. O. Google Scholar
Franco, Celia R. C. Google Scholar
Boucas, Rodrigo I. Google Scholar
Jarrouge, Thais R. Google Scholar
Pinhal, Maria A. S. Google Scholar
Tersariol, Ivarne L. S. Google Scholar
Gouvea, Tiago C. Google Scholar
Dietrich, Carl P. Google Scholar
Nader, Helena B. Autor UNIFESP Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Univ Fed Parana
Universidade de São Paulo (USP)
Univ Fed Rio Grande do Norte
Resumo Exposure of endothelial cells to heparin and other antithrombotic drugs specifically stimulates the synthesis of an antithrombotic heparan sulfate (HS). in the present work, biotinylated heparin (BiotHep) was used to characterize the binding site(s) of heparin responsible for the stimulus in HS synthesis on endothelial cells. No differences were observed between biotinylated and non-biotinylated heparin in their ability to increase the synthesis of HS. in kinetic studies the BiotHep showed fast, saturable and specific binding with an apparent K-D of 83 nM to adherent cells and 44 nM to the extracellular matrix (ECM) in the absence of cells. By confocal and electron microscopy, BiotHep bound only to the ECM co-localizing with fibronectin. the same pattern of binding to the ECM was observed using heparin conjugated with FITC or Alexa Fluor 488 in the presence or absence of fetal calf serum. However, after degradation of HS, heparin binds to the cell surface, indicating that endogenous HS possibly occupied the heparin binding sites. Analyses by flow cytometry and confocal microscopy of cells with non-associated ECM, showed labeling of the cell surface using syndecan-4 monoclonal antibody as well as wheat germ agglutinin, but no binding of heparin. Furthermore, the stimulation in HS synthesis is not elicited by heparin in the absence of ECM. These results indicate that the stimulus for the synthesis of HS does not require binding of the heparin to the cell surface, and the signaling may be mediated through the ECM.
Idioma Inglês
Financiador Financiadora de Estudos e Projetos (FINEP), Brazil
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Data de publicação 2008-11-01
Publicado em Journal of Cellular Physiology. Hoboken: Wiley-liss, v. 217, n. 2, p. 328-337, 2008.
ISSN 0021-9541 (Sherpa/Romeo, fator de impacto)
Publicador Wiley-Blackwell
Extensão 328-337
Fonte http://dx.doi.org/10.1002/jcp.21504
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000259721800006
Endereço permanente http://repositorio.unifesp.br/handle/11600/30987

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