Involvement of heparan sulfate proteoglycans in cellular uptake of high molecular weight kininogen.
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.
Melo
Katia R B
KR
Department of Biochemistry, Universidade Federal de São Paulo, Brazil.
Gutierrez
Augusto
A
Nascimento
Fabio D
FD
Araújo
Mariana Kátia Fábio S
MK
Sampaio
Misako U
MU
Carmona
Adriana K
AK
Coulson-Thomas
Yvette M
YM
Trindade
Edvaldo S
ES
Nader
Helena B
HB
Tersariol
Ivarne L S
IL
Motta
Guacyara
G
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
D019812:Heparan Sulfate Proteoglycans; D019679:Kininogen, High-Molecular-Weight; D011509:Proteoglycans