Activation of human prothrombin by human prothrombinase. Influence of factor Va on the reaction mechanism.

Krishnaswamy, Sriram; Church, William R.; Nesheim, Michael E.; Mann, Kenneth G.

doi:10.1016/s0021-9258(18)61503-0

Cited by 264 publications

(84 citation statements)

References 34 publications

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“…The multidomain assembly comprises an N-terminal Gla domain (residues 1-46), kringle 1 (residues 65-143), kringle 2 (residues 170-248) and a C-terminal protease domain (residues 285-579) connected by three intervening linkers (Figure 1A) (2)(3)(4)(5). Thrombin formation requires cleavage of prothrombin at two sites, R271 and R320, by the prothrombinase complex composed of factor Xa (fXa), cofactor Va (fVa) and phospholipids (6)(7)(8)(9). Cleavage at R271 sheds the Gla domain and two kringles and generates the inactive intermediate prethrombin-2 (Figure 1A).…”

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confidence: 99%

Role of sequence and position of the cleavage sites in prothrombin activation

Stojanovski

2021

Journal of Biological Chemistry

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confidence: 99%

Role of sequence and position of the cleavage sites in prothrombin activation

Stojanovski

2021

Journal of Biological Chemistry

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“…Furthermore, FVa appeared to cancel out the possible hinderance between FXa and prothrombin introduced by the N-glycan at K148N/R150T, indicating that the cofactor either was able to push the glycan out of the way or was inducing a conformational change in this region, hence eliminating the effect of N-glycan at this position. It has been reported previously [26, 27] that prothrombin activation can occur via two mechanistically different pathways, comprising intermediates prethrombin or meizothrombin, depending on absence or presence of phospholipids and cofactor.…”

Section: Discussionmentioning

confidence: 99%

Prolonging coagulant activity of factor Xa under hemophilic conditions by site-specific N-glycosylation of the surface-exposed autolysis loop

Bonde

Lund

Hansen

et al. 2021

Preprint

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The regulation of Factor X (FX) is critical to maintain hemostasis. To gain insights to the regulation of the active and zymogen form of coagulation FX, we probed specific molecular interactions by introducing novel N-linked glycosylations on the surface-exposed loop spanning residues 143-150 (chymotrypsin numbering) of FX. Introduction of N-glycans in the autolysis loop of these FX variants decreased Factor VIIa (FVIIa)-mediated activation ~3-fold and prothrombin activation 2- to 10-fold presumably through steric hinderance. Prothrombin activation was, however, recovered in presence of cofactor Factor Va (FVa) despite a reduced prothrombinase assembly. The introduced N-glycans exhibited position-specific effects on the interaction with two FXa inhibitors: tissue factor pathway inhibitor (TFPI) and antithrombin (ATIII). Ki for the inhibition by full-length TFPI of these FXa variants was increased by 7- to 1150-fold, while ATIII inhibition in the presence of the heparin-analogue Fondaparinux was modestly increased by 2- to 15-fold compared to wild type. To probe the in vitro hemostatic effect of the FX variants, the thrombin generation potential in FX-depleted plasma was evaluated. When supplemented in zymogen form, the FX variants exhibited reduced thrombin generation activity relative to wild-type FX, whereas enhanced procoagulant activity was measured for activated FX variants with N-glycosylation at positions 148-150. These results indicate that residues of the surface-exposed autolysis loop and residues close by participate in FX activation, proteolytic activity and inhibition of FXa by TFPI and ATIII. In plasma-based assays, a modest decrease in FX-activation rate appeared to compensate for the collective reduction in inhibitor interactions.

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“…1), as described by the groups of Krishnaswamy and Nesheim in the 1980s. 26,27 This reaction has been studied extensively, and much important information has been gained, which has enhanced our understanding of serine protease activity and coagulation factor activation in general. 28 The main activator of prothrombin is the prothrombinase complex (FXa þ FVa), which is itself activated through exposure to tissue factor (TF) FVIIa or FIX-FVIIIa as described in detail later (see also ►Fig.…”

Section: Activation Of Prothrombin To Thrombinmentioning

confidence: 99%

Thrombin: A Pivotal Player in Hemostasis and Beyond

Larsen

Hvas

2021

Semin Thromb Hemost

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The serine protease thrombin, a naturally derived enzyme, plays a key role in hemostasis by converting fibrinogen to fibrin and activating coagulation factor XIII whereby the fibrin clot is stabilized. Furthermore, thrombin activates platelets through protease-activated receptors on the platelet surface. Conversely, thrombin also exerts anticoagulant effects, enhancing the protein C activity while complexed with thrombomodulin. During recent years, it has become evident that thrombin has significant effects beyond hemostasis, as it contributes also to modulation of the endothelium, promotes inflammation and angiogenesis, and plays a role in tumor progression. Yet, due to the very short half-life and almost immediate inhibition in fluid phase by antithrombin, thrombin itself remains elusive, and only indirect measurement of thrombin generation is possible. This review provides a description of structure and mechanisms of action of thrombin both in physiological and pathological processes. Furthermore, it summarizes laboratory tests that measure in vivo or ex vivo thrombin generation, and presents knowledge on the value of these biomarkers in bleeding disorders, cardiopulmonary bypass surgery, and thromboembolic risk assessment in different patient populations. Finally, this review outlines further perspectives on using thrombin generation biomarkers for research purposes and in clinical practice.

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Activation of human prothrombin by human prothrombinase. Influence of factor Va on the reaction mechanism.

Cited by 264 publications

References 34 publications

Role of sequence and position of the cleavage sites in prothrombin activation

Role of sequence and position of the cleavage sites in prothrombin activation

Prolonging coagulant activity of factor Xa under hemophilic conditions by site-specific N-glycosylation of the surface-exposed autolysis loop

Thrombin: A Pivotal Player in Hemostasis and Beyond

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