Role of Electrostatic Interactions in Binding of Thrombin to the Fibrinogen γ′ Chain

Alexander, Kristine S.; Fried, Mike; Farrell, David H.

doi:10.1021/bi2016519

Cited by 11 publications

(11 citation statements)

References 24 publications

(80 reference statements)

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“…With the exception of WT‐S2, all peptides prolonged the lag time and decreased the peak height of thrombin generation, confirming the overall anticoagulant effect of the fibrinogen γ′ chain. Notably, tyrosine sulfation considerably increased the potency of the peptide, in line with the notion that the two tyrosine residues of the γ′ chain are sulfated in vivo and play a critical role in the interaction with thrombin exosite II . Interestingly, in our assay the doubly sulfated peptide was a more potent anticoagulant than the doubly phosphorylated peptide, which has been used in previous studies .…”

Section: Discussionsupporting

confidence: 87%

“…Although the K d for the interaction of the (doubly phosphorylated) fibrinogen c′ peptide with thrombin exosite II has been estimated to be < 1 lmol L -1 [8,30], in our and other studies [11,32] much higher peptide concentrations were needed to show appreciable effects of the peptide on coagulation in plasma. Presently, we have no explanation for this discrepancy.…”

Section: No Peptidecontrasting

confidence: 60%

“…Time-courses of FV activation were fitted to a single exponential curve and initial rates of FV activation were plotted as a function of peptide concentration (C). [3,29] and play a critical role in the interaction with thrombin exosite II [10,30]. Interestingly, in our assay the doubly sulfated peptide was a more potent anticoagulant than the doubly phosphorylated peptide, which has been used in previous studies [8,11,30,31].…”

Section: No Peptidementioning

confidence: 90%

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Inhibition of thrombin‐mediated factor V activation contributes to the anticoagulant activity of fibrinogen γ′

Omarova

Willige

Ariëns

et al. 2013

Journal of Thrombosis and Haemostasis

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To cite this article: Omarova F, Uitte De Willige S, Ari€ ens RAS, Rosing J, Bertina RM, Castoldi E. Inhibition of thrombin-mediated factor V activation contributes to the anticoagulant activity of fibrinogen c′. J Thromb Haemost 2013; 11: 1669-78.Summary. Background: Besides its role in blood clotting, fibrinogen exerts a poorly understood anticoagulant function by binding thrombin and modulating its activity. In particular, the cA/c′ fibrinogen isoform binds with high affinity to thrombin exosite II through the anionic carboxyl-terminal end of the c′ chain. This interaction downregulates thrombin-mediated factor VIII (FVIII) activation, but its effect on FV activation is unknown. Objectives: To investigate the overall anticoagulant activity of fibrinogen and particularly of fibrinogen c′ in plasma, and to verify whether the fibrinogen c′ carboxyl-terminal peptide affects thrombin-mediated FV activation. Methods: Thrombin generation was measured by calibrated automated thrombography in whole and defibrinated plasma and in plasma supplemented with the (sulfated) fibrinogen c′ carboxyl-terminal peptide (0-500 lmol L -1 ).The effect of the peptide on thrombin-mediated FV activation was studied in model systems and in plasma. Results: Total fibrinogen prolonged the lag time of thrombin generation at low tissue factor (TF) concentrations. The fibrinogen c′ peptide dose-dependently prolonged the lag time and decreased the peak height of thrombin generation at low TF, whereas a scrambled control peptide was ineffective. These effects persisted in the presence of an anti-FVIII antibody, suggesting that the peptide may also inhibit thrombin-mediated activation of FV. This was confirmed in model systems and in plasma. Conclusions: Total fibrinogen and the fibrinogen c′ peptide have an overall anticoagulant effect on thrombin generation determined at low TF. Inhibition of thrombinmediated FV activation by the fibrinogen c′ peptide is a novel mechanism of the anticoagulant activity of fibrinogen c′.

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Section: Discussionsupporting

confidence: 87%

Section: No Peptidecontrasting

confidence: 60%

Section: No Peptidementioning

confidence: 90%

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Inhibition of thrombin‐mediated factor V activation contributes to the anticoagulant activity of fibrinogen γ′

Omarova

Willige

Ariëns

et al. 2013

Journal of Thrombosis and Haemostasis

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“…8 The more abundant gA isoform has a short C-terminal tail of 4 amino acids (gA408-411, AGDV) containing a platelet-binding site, whereas the less abundant g9 isoform has an extended C-terminal tail of 20 amino acids (g9408-427, VPREHPAETEYDSLYPEDDL) containing a highaffinity binding site for thrombin exosite II. [9][10][11] Random pairing of the g chain isoforms generates 3 types of fibrinogen molecules: gA/gA (85%-92%), gA/g9 (8%-15%) and g9/g9 (,0.5%).…”

Section: Introductionmentioning

confidence: 99%

Fibrinogen γ′ increases the sensitivity to activated protein C in normal and factor V Leiden plasma

Omarova

Willige

Simioni

et al. 2014

Blood

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“…The importance of surface and active center charges of acetylcholinesterase in the electrostatic attraction of its ligands, and the influence of charge on the reactive orientation of the ligand has previously been demonstrated [6]. It was shown that electrostatics plays profound role in the thrombin-γ′ chain interactions [7]. The role of electrostatics on protein binding energy and its salt dependence was outlined in a series of other works [8–10].…”

Section: Introductionmentioning

confidence: 99%

The Role of Protonation States in Ligand-Receptor Recognition and Binding

Petukh¹,

Stefl²,

Alexov³

2013

CPD

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In this review we discuss the role of protonation states in receptor-ligand interactions, providing experimental evidences and computational predictions that complex formation may involve titratable groups with unusual pKa’s and that protonation states frequently change from unbound to bound states. These protonation changes result in proton uptake/release, which in turn causes the pH-dependence of the binding. Indeed, experimental data strongly suggests that almost any binding is pH-dependent and to be correctly modeled, the protonation states must be properly assigned prior to and after the binding. One may accurately predict the protonation states when provided with the structures of the unbound proteins and their complex; however, the modeling becomes much more complicated if the bound state has to be predicted in a docking protocol or if the structures of either bound or unbound receptor-ligand are not available. The major challenges that arise in these situations are the coupling between binding and protonation states, and the conformational changes induced by the binding and ionization states of titratable groups. In addition, any assessment of the protonation state, either before or after binding, must refer to the pH of binding, which is frequently unknown. Thus, even if the pKa’s of ionizable groups can be correctly assigned for both unbound and bound state, without knowing the experimental pH one cannot assign the corresponding protonation states, and consequently one cannot calculate the resulting proton uptake/release. It is pointed out, that while experimental pH may not be the physiological pH and binding may involve proton uptake/release, there is a tendency that the native receptor-ligand complexes have evolved toward specific either subcellular or tissue characteristic pH at which the proton uptake/release is either minimal or absent.

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Role of Electrostatic Interactions in Binding of Thrombin to the Fibrinogen γ′ Chain

Cited by 11 publications

References 24 publications

Inhibition of thrombin‐mediated factor V activation contributes to the anticoagulant activity of fibrinogen γ′

Inhibition of thrombin‐mediated factor V activation contributes to the anticoagulant activity of fibrinogen γ′

Fibrinogen γ′ increases the sensitivity to activated protein C in normal and factor V Leiden plasma

The Role of Protonation States in Ligand-Receptor Recognition and Binding

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