Characterization of the enzymatic activity of the serine protease domain of Factor VII activating protease (FSAP)

Nielsen, Nis Valentin; Roedel, Elfie; Manna, Dipankar; Etscheid, Michael; Morth, J. Preben; Kanse, Sandip M.

doi:10.1038/s41598-019-55531-x

Cited by 14 publications

(22 citation statements)

References 29 publications

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“…This indirectly implicates FSAP in the outcome of stroke in mouse models. We have recently expressed the recombinant serine protease domain (SPD) of human FSAP and confirmed that it recapitulates all, the tested, activities of the endogenous full-length protein 25 . Here, we report on the efficacy of FSAP-SPD treatment in mouse models of ischemic stroke.…”

Section: Introductionmentioning

confidence: 74%

“…WT-SPD provoked a hemostasis response, most probably, due to the inactivation of tissue factor pathway inhibitor 12,25 . The seemingly incongruent effects; the ability to promote coagulation and, at the same time, improve stroke outcome can be reconciled with the hypothesis that in the stroke model the pro-fibrinolytic effect of FSAP predominates over the pro-coagulant effect leading to an overall positive outcome.…”

Section: Resultsmentioning

confidence: 99%

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Factor VII activating protease (FSAP) inhibits the outcome of ischemic stroke in mouse models

Kim

Manna

Leergaard

et al. 2022

Preprint

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Factor VII activating protease (FSAP) is a circulating serine protease, and individuals with the Marburg I (MI) single nucleotide polymorphism (SNP), which results in an inactive enzyme, have an increased risk of stroke. The outcome of ischemic stroke is more marked in FSAP-deficient mice compared to their wild-type (WT) counterparts. Plasma FSAP levels are raised in patients as well as mice after stroke. In vitro, FSAP promotes fibrinolysis by cleavage of fibrinogen, activates protease-activated receptors and decreases the cellular cytotoxicity of histones. Since these are desirable properties in stroke treatment, we tested the effect of recombinant serine protease domain of FSAP (FSAP-SPD) on ischemic stroke in mice. A combination of tissue plasminogen activator (tPA) and FSAP-SPD enhanced clot lysis, improved microvascular perfusion and neurological outcome and reduced infarct volumes in a mouse model of thromboembolic stroke. In the tail bleeding model FSAP-SPD treatment provoked a faster clotting time indicating that it has a pro-coagulant effect that is described before. FSAP-SPD improved stroke outcome and diminished the negative effects of co-treatment with tPA in the transient middle cerebral artery occlusion model. The inactive MI-isoform of FSAP did not have any effects in either model. In mice with FSAP deficiency there were minor differences in the outcomes of stroke but the treatment with FSAP-SPD was equally effective. Thus, FSAP represents a promising novel therapeutic strategy in the treatment of ischemic stroke that requires further evaluation.

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Section: Introductionmentioning

confidence: 74%

Section: Resultsmentioning

confidence: 99%

Factor VII activating protease (FSAP) inhibits the outcome of ischemic stroke in mouse models

Kim

Manna

Leergaard

et al. 2022

Preprint

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“…14,60 In contrast, the large hydrophobic side chain of Met156 in factor VII and factor VII activating protease (an enzyme with some homology to FXII and pro-HGFA) would not be expected to stabilize an open active site conformation in the zymogen. 60,71 Indeed, Met156 may contribute to the zymogen-like behavior of the activated form of FVII (FVIIa) when it is not bound to its cofactor tissue factor. Petrovan and Ruf replaced Met156 in factor VIIa with glutamine and noted increased protease activity in the absence of tissue factor.…”

Section: Con Clus I On S and Future Cons Ider Ationsmentioning

confidence: 99%

“…Modeling of the FXII and PK protease domains based on the tPA structure suggests that Gln156 can coordinate with the Asp194 carboxylate to weakly stabilize an open active site conformation through hydrogen bonding 14,60 . In contrast, the large hydrophobic side chain of Met156 in factor VII and factor VII activating protease (an enzyme with some homology to FXII and pro‐HGFA) would not be expected to stabilize an open active site conformation in the zymogen 60,71 . Indeed, Met156 may contribute to the zymogen‐like behavior of the activated form of FVII (FVIIa) when it is not bound to its cofactor tissue factor.…”

Section: Conclusion and Future Considerationsmentioning

confidence: 99%

Proteolytic activity of contact factor zymogens

Shamanaev

Emsley

Gailani

2021

Journal of Thrombosis and Haemostasis

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Contact activation is triggered when blood is exposed to compounds or “surfaces” that promote conversion of the plasma zymogens factor XII (FXII) and prekallikrein to the active proteases FXIIa and kallikrein. FXIIa promotes blood coagulation by converting zymogen factor XI (FXI) to the protease FXIa. Contact activation appears to represent an enhancement of the propensity for FXII and prekallikrein to reciprocally activate each other by surface‐independent limited proteolysis. The nature of the activities that perpetuate this process, and that trigger contact activation, are debated. FXII and prekallikrein, like most members of the chymotrypsin/trypsin protease family, are synthesized as single polypeptides that are presumed to be in an inactive state. Internal cleavage leads to conformational changes in the protease domain that convert the enzyme active site from a closed conformation to an open conformation accessible to substrates. We observed that FXII expresses a low level of activity as a single‐chain zymogen that catalyzes prekallikrein activation in solution, as well as surface‐dependent activation of prekallikrein, FXI, and FXII (autoactivation). Prekallikrein also expresses activity that promotes cleavage of kininogen to release bradykinin, and surface‐dependent FXII activation. Modeling suggests that a glutamine residue at position 156 in the FXII and prekallikrein protease domains stabilizes an open active site conformation by forming hydrogen bonds with Asp194. The activity inherent in FXII and prekallikrein suggests a mechanism for sustaining reciprocal activation of the proteins and for initiating contact activation, and supports the premise that zymogens of some trypsin‐like enzymes are active proteases.

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“…The Marburg I (MI) single nucleotide polymorphism in the FSAPencoding gene leads to an amino acid exchange in the serine protease domain of FSAP and inactivation of the enzyme [12]. This may be due to a defect in the conversion from the zymogen to the active enzyme [13,14]. This polymorphism is a risk factor for carotid stenosis [15], stroke [16] as well as liver fibrosis [17].…”

Section: Introductionmentioning

confidence: 99%

Cellular effects of factor VII activating protease (FSAP)

Byskov

Etscheid

Kanse

2020

Thrombosis Research

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Factor VII activating protease (FSAP) is a circulating serine protease of broad specificity that is likely to be involved in many pathophysiological processes. The activation of the circulating zymogen form of FSAP by histones, released from damaged cells, underlines its roles in regulating host responses to tissue damage and inflammation. Some of the direct cellular effects of FSAP are mediated through protease-activated receptors (PARs). Knock-down of each one of the four PARs in endothelial cells indicated that PAR-1 and -3 are involved in regulating endothelial permeability in response to FSAP. Overexpression of PARs in cell lines led to the conclusion that PAR-2 and -1 were the main receptors for FSAP. Studies with synthetic peptides and receptor mutants demonstrate that FSAP cleaves PAR-1 and -2 at their canonical cleavage site. However, PAR-1 is not activated by FSAP in all cells, which may be related to other, as yet, undefined factors. Inhibition of apoptosis by FSAP is mediated through PAR-1 and was observed in neurons, astrocytes and A549 cells. FSAP also mediates cellular effects by modulating the activity of growth factors, generation of bradykinin, C5a and C3a generation or histone inactivation. These cellular effects need to be further investigated at the in vivo level.

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Characterization of the enzymatic activity of the serine protease domain of Factor VII activating protease (FSAP)

Cited by 14 publications

References 29 publications

Factor VII activating protease (FSAP) inhibits the outcome of ischemic stroke in mouse models

Factor VII activating protease (FSAP) inhibits the outcome of ischemic stroke in mouse models

Proteolytic activity of contact factor zymogens

Cellular effects of factor VII activating protease (FSAP)

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