Examination of the substrate specificity of heparin and heparan sulfate lyases

Linhardt, Robert J.; Turnbull, Jeremy E.; Wang, H M; Loganathan, Duraikkannu; Gallagher, John T.

doi:10.1021/bi00462a026

Cited by 271 publications

(192 citation statements)

References 19 publications

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“…This CXCL8 binding profile and its specificity have already been described under identical binding conditions in our previous study (41). In order to investigate whether the binding involved HS, sections were treated with and without heparinases I and III (48). Sections were then incubated with 125 I-CXCL8, and gamma counting was performed.…”

Section: Resultsmentioning

confidence: 60%

Induction of a CXCL8 binding site on endothelial syndecan‐3 in rheumatoid synovium

Patterson

Gardner

Shaw

et al. 2005

Arthritis & Rheumatism

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Section: Resultsmentioning

confidence: 60%

Induction of a CXCL8 binding site on endothelial syndecan‐3 in rheumatoid synovium

Patterson

Gardner

Shaw

et al. 2005

Arthritis & Rheumatism

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“…The cells were incubated with 1 unit/ml of either heparinase I, heparinase II, or heparinase III (Sigma) for 4 h at 37°C (30). The cells were washed with starving medium twice before they were treated with antithrombin and FGF-2.…”

Section: Effect Of Heparinases On Antithrombin Inhibition Of Huvec Prmentioning

confidence: 99%

Antiangiogenic Antithrombin Blocks the Heparan Sulfate-dependent Binding of Proangiogenic Growth Factors to Their Endothelial Cell Receptors

Zhang

Swanson

Xiong

et al. 2006

Journal of Biological Chemistry

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The anticoagulant serpin antithrombin acquires a potent antiangiogenic activity upon undergoing conformational alterations to cleaved or latent forms. Here we show that antithrombin antiangiogenic activity is mediated at least in part through the ability of the conformationally altered serpin to block the proangiogenic growth factors fibroblast growth factor (FGF)-2 and vascular endothelial growth factor (VEGF) from forming signaling competent ternary complexes with their protein receptors and heparan sulfate co-receptors on endothelial cells. Cleaved and latent but not native forms of antithrombin blocked the formation of FGF-2-FGF receptor-1 ectodomain-heparin ternary complexes, and the dimerization of these complexes in solution and similarly inhibited the formation of FGF-2-heparin binary complexes and their dimerization. Only antiangiogenic forms of antithrombin likewise inhibited125 I-FGF-2 binding to its low affinity heparan sulfate co-receptor and blocked FGF receptor-1 autophosphorylation and p42/44 MAP kinase phosphorylation in cultured human umbilical vein endothelial cells (HUVECs). Moreover, treatment of HUVECs with heparinase III to specifically eliminate the FGF-2 heparan sulfate co-receptor suppressed the ability of antiangiogenic antithrombin to inhibit growth factor-stimulated proliferation. Antiangiogenic antithrombin inhibited full-length VEGF 165 stimulation of HUVEC proliferation but did not affect the stimulation of cells by the heparin-binding domain-deleted VEGF 121 . Taken together, these results demonstrate that antiangiogenic forms of antithrombin block the proangiogenic effects of FGF-2 and VEGF on endothelial cells by competing with the growth factors for binding the heparan sulfate co-receptor, which mediates growth factor-receptor interactions. Moreover, the inability of native antithrombin to bind this co-receptor implies that native and conformationally altered forms of antithrombin differentially bind proangiogenic heparan sulfate domains.Angiogenesis, the growth of new capillaries from pre-existing vessels, is a key physiologic process whose dysregulation underlies many diseases (1). It is particularly important for the transition of tumors from a dormant state to a malignant state, because new vessels supply oxygen, essential nutrients, and growth factors that allow the tumor mass to expand. This expectation has led to the idea that angiogenesis inhibitors may be useful antitumor drugs (2). A number of naturally occurring angiogenesis inhibitors have been identified as potential antitumor therapeutic agents that are derived by modification of endogenous proteins, e.g. angiostatin is a fragment of plasminogen (3), endostatin is a fragment produced by proteolytic cleavage of collagen XVIII (4), and the serpin antithrombin acquires antiangiogenic activity upon undergoing conformational alterations induced by mild heating or protease cleavage (5).Interestingly, whereas a number of serpins, such as maspin, pigment epithelium-derived factor, and kallistatin, have been shown to pos...

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“…When the HUVEC monolayers are treated with TGF- 1 , HSPG synthesis is stimulated and the amount of HSPGs on the surface of the HUVEC monolayer increases [37,38]. This results in an increase in the amount of glycocalyx, either by increase in the number or in the length of the brushes.…”

Section: Discussion Of Ecs Gel Friction In Vitro Using Hydrogel Lubrimentioning

confidence: 99%

“…On the other hand, to disrupt HSPGs, HUVEC monolayers were incubated for 4 h in protein-free HEM containing 1.45 U/ml heparinase I (Sigma) [17,38]. Detection of HSPGs was performed as follows [39]:…”

Section: Glycocalyx Modulation and Immunofluorescencementioning

confidence: 99%

“…Of proteoglycans, the main component is heparan sulfate proteoglycans (HSPGs) (5090wt%), which consist of a core protein and heparan sulfate-type glycosaminoglycan, containing numerous negatively charged sulfate groups and binds several plasma proteins [7]. The amount of glycocalyx on the EC surface can be enhanced using TGF- 1 , a cytokine that stimulate heparan sulfate proteoglycans (HSPGs), the main proteoglycan of glycocalyx synthesis [37,38]. It was reported that TGF- 1 treatment can increase the HSPG content on the EC surface by more than 50%.…”

Section: Effect Of Glycocalyx On Ec Monolayer Friction 321 Glycocalmentioning

confidence: 99%

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Study on the Sliding Friction of Endothelial Cells Cultured on Hydrogel and the Role of Glycocalyx on Friction Reduction

et al. 2010

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In this study, we investigated the sliding friction of human umbilical vein endothelial cell (HUVEC) monolayer cultured on poly(sodium p-styrene sulfonate) (PNaSS) gel, intending to elucidate the role of the glycocalyx on the surface of endothelial cell (EC) in friction reduction.Three sets of HUVEC monolayers were investigated: 1) as-cultured HUVEC monolayer, 2) HUVEC monolayer treated by transforming growth factor  1 (TGF- 1 ), which increased glycocalyx by 148%, 3) HUVEC monolayer treated by heparinase I, which reduced glycocalyx by 57%, both were compared with that of the as prepared one. When being slid on flat glass surface, the frictional stress of HUVEC monolayer decreased in the order of heparinase I-treated > as-cultured > TGF- 1 -treated samples. The results suggested that glycocalyx may play a role in reducing the friction of endothelial cell monolayer.

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Examination of the substrate specificity of heparin and heparan sulfate lyases

Cited by 271 publications

References 19 publications

Induction of a CXCL8 binding site on endothelial syndecan‐3 in rheumatoid synovium

Induction of a CXCL8 binding site on endothelial syndecan‐3 in rheumatoid synovium

Antiangiogenic Antithrombin Blocks the Heparan Sulfate-dependent Binding of Proangiogenic Growth Factors to Their Endothelial Cell Receptors

Study on the Sliding Friction of Endothelial Cells Cultured on Hydrogel and the Role of Glycocalyx on Friction Reduction

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