The functional properties of a truncated form of endothelial cell protein C receptor generated by alternative splicing

Molina, Eva; Hermida, José; López‐Sagaseta, Jacinto; Puy, Cristina; Montes, Ramón

doi:10.3324/haematol.12376

Cited by 10 publications

(5 citation statements)

References 21 publications

(35 reference statements)

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“…Thus, the existence of two distinct soluble species of NRP-1 containing either all extracellular domains (120 kDa) (Lu et al, 2009; Swendeman et al, 2008) or the four of shNRP-1 (75 kDa) (Xu et al, 2008) may have distinct functions; the latter would not be retained on the HS of the shedding cell’s pericellular matrix. Differences between membrane bound and soluble, shed species have been observed for other proteins, e.g., receptor for advanced glycation products (RAGE) and endothelial cell protein C receptor (EPCR) (Koyama, Yamamoto & Nishizawa, 2007; Molina et al, 2008). Thus, the different heparin-binding properties of the dimerised/oligomeric NRP-1 and the monomeric isoforms might drive distinct output responses of the cells.…”

Section: Resultsmentioning

confidence: 99%

Characterisation of the interaction of neuropilin-1 with heparin and a heparan sulfate mimetic library of heparin-derived sugars

Uniewicz

Ori

Ahmed

et al. 2014

PeerJ

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Background. Neuropilin-1 (NRP-1) is a multidomain membrane protein with soluble isoforms interacting with a complex network of other membrane receptors, their respective ligands and heparan sulfate (HS). It is involved in the development of vasculature, neural patterning, immunological responses and pathological angiogenesis.Methods. We have characterised the binding of a Fc fusion of rat NRP-1 (Fc rNRP-1) and of a soluble isoform, corresponding to the first four extracellular domains of human NRP-1, shNRP-1, using optical biosensor-based binding assays with a library of heparin derivatives. Selective labelling of lysines protected upon heparin binding allowed their identification by mass spectrometry.Results. Fc rNRP-1 bound to heparin with high affinity (2.5 nM) and fast ka (9.8 × 106 M−1s−1). Unusually, NRP-1 bound both highly sulfated and completely desulfated stretches of heparin and exhibited a complex pattern of preferences for chemically modified heparins possessing one or two sulfate groups, e.g., it bound heparin with just a 6-O sulfate group better than heparin with any two of N-sulfate, 6-O sulfate and 2-O sulfate. Mass-spectrometry based mapping identified that, in addition to the expected the b1 domain, the a1, and c domains and the L2 linker were also involved in the interaction. In contrast, shNRP-1 bound heparin far more weakly. This could only be shown by affinity chromatography and by differential scanning fluorimetry.Discussion. The results suggest that the interaction of NRP-1 with HS is more complex than anticipated and involving a far greater extent of the protein than just the b1–b2 domains. NRP-1’s preference for binding long saccharide structures suggests it has the potential to bind large segments of HS chains and so organise their local structure. In contrast, the four domain soluble isoform, shNRP-1 binds heparin weakly and so would be expected to diffuse away rapidly from the source cell.

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Section: Resultsmentioning

confidence: 99%

Characterisation of the interaction of neuropilin-1 with heparin and a heparan sulfate mimetic library of heparin-derived sugars

Uniewicz

Ori

Ahmed

et al. 2014

PeerJ

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“…This substitution has been shown to result in increased sensitivity to EPCR cell surface shedding by ADAM17 [60]. Recent evidence indicates that an additional means by which the A3 haplotype contributes to elevated sEPCR levels is via expression of an alternatively spliced truncated PROCR mRNA [61,62]. This alternatively spliced mRNA isoform is truncated at the 3 0 end of exon 3, and therefore does not encode exon 4.…”

Section: Regulation Of Coagulation By Epcr-ligand Interactionsmentioning

confidence: 99%

The endothelial cell protein C receptor: cell surface conductor of cytoprotective coagulation factor signaling

Gleeson

O’Donnell

Preston

2011

Cell. Mol. Life Sci.

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Increasing evidence links blood coagulation proteins with the regulation of acute and chronic inflammatory disease. Of particular interest are vitamin K-dependent proteases, which are generated as a hemostatic response to vascular injury, but can also initiate signal transduction via interactions with vascular receptors. The endothelial cell protein C receptor (EPCR) is a multi-ligand vitamin K-dependent protein receptor for zymogen and activated forms of plasma protein C and factor VII. Although the physiological role of the EPCR-FVII(a) interaction is not well-understood, protein C binding to EPCR facilitates rapid generation of APC in response to excessive thrombin generation, and is a central requirement for the multiple signal-transduction cascades initiated by APC on both vascular endothelial and innate immune cells. Exciting recent studies have highlighted the emerging role of EPCR in modulating the cytoprotective properties of APC in a number of diverse inflammatory disorders. In this review, we describe the structure-function relationships, signal transduction pathways, and cellular interactions that enable EPCR to modulate the anticoagulant and anti-inflammatory properties of its vitamin K-dependent protein ligands, and examine the relevance of EPCR to both thrombotic and inflammation-associated disease.

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“…In this issue of the journal, Molina and co-workers 35 report the production of a truncated EPCR mRNA by HUVECs, similar to the isoform we described. More interestingly, they showed that the corresponding protein was produced and secreted by cultured lung cancer cells.…”

Section: Acquired Epcr Abnormalities and Increased Sepcr Levelsmentioning

confidence: 98%

Endothelial cell protein C receptor and the risk of venous thrombosis

Gandrille¹

2008

Haematologica

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The functional properties of a truncated form of endothelial cell protein C receptor generated by alternative splicing

Cited by 10 publications

References 21 publications

Characterisation of the interaction of neuropilin-1 with heparin and a heparan sulfate mimetic library of heparin-derived sugars

Characterisation of the interaction of neuropilin-1 with heparin and a heparan sulfate mimetic library of heparin-derived sugars

The endothelial cell protein C receptor: cell surface conductor of cytoprotective coagulation factor signaling

Endothelial cell protein C receptor and the risk of venous thrombosis

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