Coagulation Factor X Interaction with Macrophages through Its N-Glycans Protects It from a Rapid Clearance

Kurdi, Mohamad; Cherel, Ghislaine; Lenting, Peter J.; Denis, Cécile V.; Olivier, Christophe

doi:10.1371/journal.pone.0045111

Cited by 10 publications

(25 citation statements)

References 33 publications

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“…Previously, we have shown that FX is targeted to resident liver macrophages, and that GdCl 3 -mediated macrophage inactivation severely reduces FX plasma levels. 16,20 Combined with the observation that FX remains at the surface of macrophages, rather than being internalized and degraded, 16,20 these data point to macrophages serving a protective role in maintaining FX plasma levels. To better understand this protective role, we searched for receptors that are involved in the binding of FX to the macrophage surface.…”

Section: Discussionmentioning

confidence: 96%

“…In a previous study conducted in our laboratory, organ biodistribution analysis identified the liver as a major target organ for FX. 16 At the cellular level, we demonstrated that FX binds to Kupffer cells, the tissue-resident macrophages of the liver. Strikingly, we observed that macrophage inactivation in mice significantly reduced FX plasma levels.…”

Section: Introductionmentioning

confidence: 82%

“…Moreover, no binding of deglycosylated FX to macrophage-expressed SR-AI could be detected in this Duolink-PLA analysis (not shown), compatible with our previous finding that N-linked glycans on FX are needed for binding to macrophages. 16 To confirm the close proximity between FX and SR-AI, classic double-staining experiments were performed using THP1 macrophages as well as stably transfected cells expressing SR-AI. In both cases, an apparent colocalization was observed when cells were analyzed via confocal microscopy ( Figures 2G and 4J).…”

Section: Discussionmentioning

confidence: 99%

“…16 To explore the specificity of FX binding to macrophages vs monocytic cells, we compared binding of Alexa 488-labeled FX to undifferentiated THP1 cells and THP1-derived macrophages. Following incubation (1 hour at 37°C), flow cytometry analysis revealed a similar amount of fluorescence for undifferentiated THP1 cells incubated either with control Alexa 488-conjugated Fab92 or Alexa 488-labeled FX (1.5 6 1.0 3 10 3 and 3.2 6 1.9 3 10 3 fluorescence units; P .…”

Section: Fx Binds To Differentiated Macrophages But Not To Monocytesmentioning

confidence: 99%

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Macrophage receptor SR-AI is crucial to maintain normal plasma levels of coagulation factor X

Muczynski

Bazaa

Loubière

et al. 2016

Blood

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Key Points• SR-AI is the major receptor of FX at the macrophage surface.• Macrophages use SR-AI to control FX circulatory levels.Beside its classical role in the coagulation cascade, coagulation factor X (FX) is involved in several major biological processes including inflammation and enhancement of virusinduced immune responses. We recently reported that the long circulatory half-life of FX is linked to its interaction with liver-resident macrophages. Importantly, we now observed that macrophages, but not undifferentiated monocytes, support this interaction. Using cell biology approaches with primary and THP1-derived macrophages as well as transfected cells, we further identified the scavenger receptor type A member I (SR-AI) to be a macrophage-specific receptor for FX. This result was confirmed using SR-AI-deficient mice, which exhibit reduced circulating levels of FX in vivo and loss of FX-macrophage interactions in vitro. Binding studies using purified proteins revealed that FX binds specifically (half-maximal binding, 3 mg/mL) to the extracellular domain of SR-AI. Altogether, we demonstrate that macrophages regulate FX plasma levels in an SR-AI-dependent manner. (Blood. 2016;127(6):778-786)

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Section: Discussionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Section: Fx Binds To Differentiated Macrophages But Not To Monocytesmentioning

confidence: 99%

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Macrophage receptor SR-AI is crucial to maintain normal plasma levels of coagulation factor X

Muczynski

Bazaa

Loubière

et al. 2016

Blood

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“…Interestingly, recent studies have reported that N-linked glycan expression on factor X also plays a key role in regulating interaction with macrophages. 59 Although the molecular mechanism through which N-linked glycan determinants regulate coagulation glycoprotein clearance remains unknown, the effect may be due to general properties of the complex sugar chains or instead may be attributable to particular carbohydrate structures located at specific N-linked sites. 42,43 Given the role of the A1-A2-A3 domains in modulating the interaction of VWF with macrophages, we further investigated whether the 2 N-linked glycosylation sites located at N1515 and N1574, respectively, might play a particular role in regulating VWF clearance.…”

Section: Discussionmentioning

confidence: 99%

N-linked glycans within the A2 domain of von Willebrand factor modulate macrophage-mediated clearance

Chion

O’Sullivan

Drakeford

et al. 2016

Blood

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Key Points• The A1 domain of VWF contains a cryptic binding site that plays a key role in regulating macrophage binding and clearance.• The N-linked glycans presented at N1515 and N1574 within the A2 domain of VWF modulate macrophage-mediated clearance.Enhanced von Willebrand factor (VWF) clearance is important in the etiology of von Willebrand disease. However, the molecular mechanisms underlying VWF clearance remain poorly understood. In this study, we investigated the role of VWF domains and specific glycan moieties in regulating in vivo clearance. Our findings demonstrate that the A1 domain of VWF contains a receptor-recognition site that plays a key role in regulating the interaction of VWF with macrophages. In A1-A2-A3 and full-length VWF, this macrophage-binding site is cryptic but becomes exposed following exposure to shear or ristocetin. Previous studies have demonstrated that the N-linked glycans within the A2 domain play an important role in modulating susceptibility to ADAMTS13 proteolysis. We further demonstrate that these glycans presented at N1515 and N1574 also play a critical role in protecting VWF against macrophage binding and clearance. Indeed, loss of the N-glycan at N1515 resulted in markedly enhanced VWF clearance that was significantly faster than that observed with any previously described VWF mutations. In addition, A1-A2-A3 fragments containing the N1515Q or N1574Q substitutions also demonstrated significantly enhanced clearance. Importantly, clodronate-induced macrophage depletion significantly attenuated the increased clearance observed with N1515Q and N1574Q in both full-length VWF and A1-A2-A3.Finally, we further demonstrate that loss of these N-linked glycans does not enhance clearance in VWF in the presence of a structurally constrained A2 domain. Collectively, these novel findings support the hypothesis that conformation of the VWF A domains plays a critical role in modulating macrophage-mediated clearance of VWF in vivo. (Blood. 2016;128(15):1959-1968

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Site‐specific functional roles of the Factor X activation peptide in the intrinsic tenase‐mediated Factor X activation

et al. 2022

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The conversion of zymogen Factor X (FX) to an active protease involves the removal of a 52‐residue long activation peptide (AP). Through site‐directed mutagenesis, we investigate the role of the AP and demonstrate that the high abundance of proline residues is important for efficient proteolysis of FX. Moreover, we identify an essential interaction site for Factor IXa (FIXa) between residues 22 and 30 (AP numbering) and find that the residues between 31 and 41 may provide an important interaction site for the intrinsic tenase complex, composed of Factor IXa (FIXa) and Factor VIIIa (FVIIIa). Finally, we suggest that the carbohydrate chain at Asn‐39 restricts the activator specificity, as elimination of this glycosylation site increases the activation rate for activation by FIXa and FXa.

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Coagulation Factor X Interaction with Macrophages through Its N-Glycans Protects It from a Rapid Clearance

Cited by 10 publications

References 33 publications

Macrophage receptor SR-AI is crucial to maintain normal plasma levels of coagulation factor X

Macrophage receptor SR-AI is crucial to maintain normal plasma levels of coagulation factor X

N-linked glycans within the A2 domain of von Willebrand factor modulate macrophage-mediated clearance

Site‐specific functional roles of the Factor X activation peptide in the intrinsic tenase‐mediated Factor X activation

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