Factor XII binding to endothelial cells depends on caveolae

Schousboe, Inger; Thomsen, Peter; Deurs, Bo van

doi:10.1111/j.1432-1033.2004.04229.x

Cited by 9 publications

(7 citation statements)

References 25 publications

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“…In our previous work we demonstrated that in CHO-K1 pretreated with methyl-β-cyclodextrin H-kininogen endocytosis is prevented and it did not colocalize with transferrin in the early phase of the endocytic process strongly indicating that the endocytosis is not dependent on clathrin [ 17 ]. Consistent with our findings, FXII binding to HUVECs depends on intact caveolae on the cellular surface [ 31 ], and caveolae harbor proteoglycans [ 32 ]. The bradykinin-free H-kininogen does not internalize as shown in the present work ( Fig.…”

Section: Discussionsupporting

confidence: 90%

Bradykinin Release Avoids High Molecular Weight Kininogen Endocytosis

et al. 2015

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Human H-kininogen (120 kDa) plays a role in many pathophysiological processes and interacts with the cell surface through protein receptors and proteoglycans, which mediate H-kininogen endocytosis. In the present work we demonstrate that H-kininogen containing bradykinin domain is internalized and different endogenous kininogenases are present in CHO-K1 cells. We used CHO-K1 (wild type) and CHO-745 (mutant deficient in proteoglycans biosynthesis) cell lines. H-kininogen endocytosis was studied using confocal microscopy, and its hydrolysis by cell lysate fraction was determined by immunoblotting. Bradykinin release was also measured by radioimmunoassay. H-kininogen interaction with the cell surface of CHO-745 cells resulted in bradykinin release by serine proteases. In CHO-K1 cells, which produce heparan and chondroitin sulfate proteoglycans, internalization of H-kininogen through its bradykinin domain can occur on lipid raft domains/caveolae. Nevertheless bradykinin-free H-kininogen was not internalized by CHO-K1 cells. The H-kininogen present in acidic endosomal vesicles in CHO-K1 was approximately 10-fold higher than the levels in CHO-745. CHO-K1 lysate fractions were assayed at pH 5.5 and intact H-kininogen was totally hydrolyzed into a 62 kDa fragment. By contrast, at an assay pH 7.4, the remained fragments were 115 kDa, 83 kDa, 62 kDa and 48 kDa in size. The antipain-Sepharose chromatography separated endogenous kininogenases from CHO-K1 lysate fraction. No difference was detected in the assays at pH 5.5 or 7.4, but the proteins in the fraction bound to the resin released bradykinin from H-kininogen. However, the proteins in the unbound fraction cleaved intact H-kininogen at other sites but did not release bradykinin. H-kininogen can interact with extravascular cells, and is internalized dependent on its bradykinin domain and cell surface proteoglycans. After internalization, H-kininogen is proteolytically processed by intracellular kininogenases. The present data also demonstrates that serine or cysteine proteases in lipid raft domains/caveolae on the CHO cell can hydrolyze H-kininogen, thus releasing kinins.

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Section: Discussionsupporting

confidence: 90%

Bradykinin Release Avoids High Molecular Weight Kininogen Endocytosis

et al. 2015

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“…It has been shown that FXII can bind to ECs in a zinc-dependent manner via urokinase-like plasminogen activator receptor (u-PAR), cytokeratin 1 and the complement receptor gC1qR1 (14,31,32). ECs are also able to produce an extracellular matrix protein which can serve as a zinc-independent binding site for FXII (13).…”

Section: Discussionmentioning

confidence: 99%

Coagulation on Endothelial Cells: The Underexposed Part of Virchow’s Triad

Geenen

Post²,

Molin³

et al. 2012

Thromb Haemost

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The process of thrombin generation involves numerous plasma proteases and cofactors. Interaction with the vessel wall, in particular endothelial cells (ECs), influences this process but data on this interaction is limited. We evaluated thrombin generation on EA.hy926, human coronary arterial ECs (HCAECs) and patient-derived human venous ECs (HVECs) by means of a modified calibrated automated thrombogram (CAT) method and especially looked into contribution of the intrinsic and extrinsic pathways. Thrombin generation was measured in presence of confluent ECs with normal pooled and factor XII-deficient (FXII-deficient) platelet-poor plasma, with/without active site inhibited factor VIIa (ASIS) to block the extrinsic pathway and corn trypsin inhibitor for blocking contact activation (intrinsic pathway). Fetal bovine serum (FBS) was removed from culture conditions as FXIIa from the serum retained on ECs apparently, thereby inducing strong contact activation. In serum-free conditions, EA.hy926 and patient-derived HVECs induced thrombin generation mainly via the contact activation pathway with minor influence of ASIS on peak height and very low thrombin generation curves in FXII-deficient plasma. HVECs derived from coronary arterial bypass graft (CABG) patients showed increased thrombin generation compared to control patients, which could be ascribed to increased contact activation. Contribution of the extrinsic pathway on patient-derived ECs was limited. We conclude that the CAT method in combination with serum-free cultured ECs offers a valuable high-throughput method to evaluate endothelial influences on thrombin generation, which appears to involve predominantly contact activation on ECs. Contact activation-mediated thrombin generation was increased on ECs from CABG patients compared to controls.

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“…HK and/or HKa binds to CK-1 through D3 and/or D5, while uPAR and gC1qR bind primarily to D5 (Mahdi et al, 2004). Interestingly, gC1qR is also a binding site for factor XII (Joseph et al, 1996) whereby binding depends on caveolae on endothelial cells and caveolae harbor GPI-anchored PGs (glypicans) (Schousboe et al, 2004). Interestingly, gC1qR is also a binding site for factor XII (Joseph et al, 1996) whereby binding depends on caveolae on endothelial cells and caveolae harbor GPI-anchored PGs (glypicans) (Schousboe et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

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

Melo

Gutierrez

Nascimento

et al. 2008

Biological Chemistry

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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.

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Factor XII binding to endothelial cells depends on caveolae

Cited by 9 publications

References 25 publications

Bradykinin Release Avoids High Molecular Weight Kininogen Endocytosis

Bradykinin Release Avoids High Molecular Weight Kininogen Endocytosis

Coagulation on Endothelial Cells: The Underexposed Part of Virchow’s Triad

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

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