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Title: Involvement of heparan sulfate proteoglycans in cellular uptake of high molecular weight kininogen
Authors: Melo, Katia R. B. [UNIFESP]
Gutierrez, Augusto [UNIFESP]
Nascimento, Fabio D. [UNIFESP]
Araujo, Mariana S. [UNIFESP]
Sampaio, Misako U. [UNIFESP]
Carmona, Adriana K. [UNIFESP]
Coulson-Thomas, Yvette May [UNIFESP]
Trindade, Edvaldo S. [UNIFESP]
Nader, Helena B. [UNIFESP]
Tersariol, Ivarne L. S. [UNIFESP]
Motta, Guacyara [UNIFESP]
Universidade Federal de São Paulo (UNIFESP)
Univ Fed Parana
Univ Mogi das Cruzes
Keywords: endocytosis
endothelial cells
kinin release
tumor cells
Issue Date: 1-Feb-2009
Publisher: Walter de Gruyter & Co
Citation: Biological Chemistry. Berlin: Walter de Gruyter & Co, v. 390, n. 2, p. 145-155, 2009.
Abstract: 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-beta-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.
ISSN: 1431-6730
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