title: Involvement of heparan sulfate proteoglycans in cellular uptake of high molecular weight kininogen.
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.
authors.affiliation: Department of Biochemistry, Universidade Federal de São Paulo, Brazil.
authors.forename: Katia R B
authors.initials: KR
authors.lastname: Melo
authors.forename: Augusto
authors.initials: A
authors.lastname: Gutierrez
authors.forename: Fabio D
authors.initials: FD
authors.lastname: Nascimento
authors.forename: Mariana Kátia Fábio S
authors.initials: MK
authors.lastname: Araújo
authors.forename: Misako U
authors.initials: MU
authors.lastname: Sampaio
authors.forename: Adriana K
authors.initials: AK
authors.lastname: Carmona
authors.forename: Yvette M
authors.initials: YM
authors.lastname: Coulson-Thomas
authors.forename: Edvaldo S
authors.initials: ES
authors.lastname: Trindade
authors.forename: Helena B
authors.initials: HB
authors.lastname: Nader
authors.forename: Ivarne L S
authors.initials: IL
authors.lastname: Tersariol
authors.forename: Guacyara
authors.initials: G
authors.lastname: Motta
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology / Q000378:metabolism; D016466:CHO Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D019812:Heparan Sulfate Proteoglycans / Q000494:pharmacology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D019679:Kininogen, High-Molecular-Weight / Q000378:metabolism*; D011509:Proteoglycans; D011817:Rabbits
chemical_list: D019812:Heparan Sulfate Proteoglycans; D019679:Kininogen, High-Molecular-Weight; D011509:Proteoglycans