Involvement of heparan sulfate proteoglycans in cellular uptake of high molecular weight kininogen

Autor(es):Mostrar menos -

Melo, Katia R. B. ^[1] ; Gutierrez, Augusto ^[1] ; Nascimento, Fabio D. ^[1] ; Araujo, Mariana S. ^[1] ; Sampaio, Misako U. ^[1] ; Carmona, Adriana K. ^[2] ; Coulson-Thomas, Yvette M. ^[1] ; Trindade, Edvaldo S. ^{[1, 3]} ; Nader, Helena B. ^[1] ; Tersariol, Ivarne L. S. ^{[1, 4]} ; Motta, Guacyara ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, UNIFESP, Dept Biochem, BR-04044020 Sao Paulo - Brazil

^[2] Univ Fed Sao Paulo, UNIFESP, Dept Biophys, BR-04044020 Sao Paulo - Brazil

^[3] Univ Fed Parana, Ctr Politecn, Dept Cell Biol, BR-81531980 Curitiba, Parana - Brazil

^[4] Univ Mogi das Cruzes, Ctr Interdisciplinar Invest Bioquim, BR-08780210 Mogi Das Cruzes, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Biological Chemistry; v. 390, n. 2, p. 145-155, 2009.
• Citações Web of Science: 7

Resumo

In this study, we analyzed the influence of proteoglycans on the interaction between human high molecular weight kininogen (HK) and the cell surface. We found that D5-related peptide inhibits HK-biotin cellular uptake. Confocal microscopy showed that HK colocalizes with heparan sulfate proteoglycan (HSPG) at the cell surface. When biotin-HK is incubated with rabbit aorta endothelial cells (RAECs) and CHO-K1 cells, it is internalized into acidic intracellular vesicles, whereas when incubated with CHO-745 cells, which express reduced levels of glycosaminoglycans, HK is not internalized. To further verify the hypothesis that HSPG-dependent mechanisms are involved in HK uptake and proteolytic processing in lysosomes, we tested chloroquine, which blocks Alexa 488-HK colocalization with Lyso Tracker in acidic endosomal vesicles. The process of HK internalization was blocked by low temperatures, methyl-β-cyclodextrin, FCCP and 2-deoxy-d-glucose, implying that HK uptake into acidic vesicles is energy-dependent and most likely involves binding to HSPG structures localized in cholesterol-rich domains present in the plasma membrane. Kinin generation at the cell surface was much higher in tumorigenic cells (CHO-K1) when compared to endothelial cells (RAECs). The present data indicate that the process of HK endocytosis involving HSPG is a novel additional mechanism which may control kinin generation at the cell surface. (AU)

• Processo FAPESP: 02/09808-6 - Estudo do processamento enzimático de proteínas plasmáticas envolvidas nos mecanismos hemostáticos
• Beneficiário: Guacyara da Motta
• Modalidade de apoio: Auxílio à Pesquisa - Regular