| Full text | |
| Author(s): Show less - |
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]
Total Authors: 11
|
| Affiliation: | [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
Total Affiliations: 4
|
| Document type: | Journal article |
| Source: | Biological Chemistry; v. 390, n. 2, p. 145-155, 2009. |
| Web of Science Citations: | 7 |
| 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-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) | |