Molecular and functional characterization of a natural homozygous Arg67His mutation in the prothrombin gene of a patient with a severe procoagulant defect contrasting with a mild hemorrhagic phenotype

Akhavan, Sepideh; Cristofaro, Raimondo De; Peyvandi, Flora; Lavoretano, Silvia; Landolfi, Raffaele; Mannucci, Pier Mannuccio

doi:10.1182/blood-2002-01-0243

Cited by 29 publications

(33 citation statements)

References 37 publications

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“…The role of proexosite I in substrate recognition is supported further by recent site-directed mutagenesis studies demonstrating that mutation of proexosite I residues in prethrombin 1 (Pre 1) results in loss of factor Va cofactor activity and is correlated with loss of affinity of the proexosite for Hir 54 -65 (SO 3 Ϫ ) (15). A natural mutation of Arg 67 to His in proexosite I of Pro isolated from a patient with a severe procoagulant defect and mild bleeding phenotype also showed reduced factor Va acceleration of its activation (16). In the preceding paper, proexosite I on Pre 1, a Pro analog that lacks the fragment 1 (F1) domain, and Pre 2 was shown to be activated by cleavage of Arg 320 -Ile 321 , generating the active intermediate, meizothrombin des-fragment 1 (MzT(-F1)), and the product, thrombin, respectively.…”

mentioning

confidence: 81%

Role of Prothrombin Fragment 1 in the Pathway of Regulatory Exosite I Formation during Conversion of Human Prothrombin to Thrombin

Anderson¹,

Bock

2003

Journal of Biological Chemistry

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mentioning

confidence: 81%

Role of Prothrombin Fragment 1 in the Pathway of Regulatory Exosite I Formation during Conversion of Human Prothrombin to Thrombin

Anderson¹,

Bock

2003

Journal of Biological Chemistry

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“…For example, reagents that block proexosite I such as hirugen and bothrojaracin inhibit macromolecular substrate cleavage by prothrombinase or the FXa-FVa complex (3,20,21). Additionally, proexosite I prethrombin-1 mutants and a naturally occurring prothrombin variant (Arg 67 to His, chymotrypsin numbering system (22)) are poor substrates for prothrombinase and FXa-FVa (23,24). Interestingly, these probes or mutations reduced the rate of IIa generation only in the presence of the cofactor, suggesting a link between FVa and proexosite I.…”

mentioning

confidence: 99%

Role of Hirudin-like Factor Va Heavy Chain Sequences in Prothrombinase Function

Toso

Camire

2006

Journal of Biological Chemistry

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Proexosite I on prothrombin has been implicated in providing a recognition site for factor Va within prothrombinase. To examine whether hirudin-like sequences (659 -698) on the cofactor contribute to this interaction, we expressed and purified two-chain FVa derivatives that were intracellularly truncated at the C terminus of the heavy chain: Blood coagulation factor Va (FVa) 2 is a heterodimeric protein composed of a heavy chain (residues 1-709; 105 kDa) and a light chain (residues 1546 -2196; 74 kDa) which arises from limited proteolysis of the pro-cofactor factor V (FV) (1). Factor Va reversibly associates with the serine protease factor Xa (FXa) on an appropriate membrane surface in the presence of calcium ions to form prothrombinase. This enzyme complex cleaves two peptide bonds in prothrombin, resulting in the generation of ␣-thrombin (IIa) (2). This reaction is also catalyzed by membrane-bound FXa; however, incorporation of FVa within prothrombinase has a profound effect on the rate of IIa generation. Additionally, prothrombinase has remarkable specificity, as its only known biological substrate is prothrombin (2).Although the molecular basis underlying the rate-enhancing effect of FVa remains largely unknown, recent contributions have provided insight into how prothrombinase recognizes its protein substrate (3-5). These studies support a model in which binding specificity is determined by two resolvable steps involving an interaction at an exosite followed by active site docking. These exosites have yet to be fully defined, but there is some evidence that at least part of the enzyme exosite lies on the catalytic domain of FXa (6 -8). Because it is well established that prothrombin binds the cofactor (9 -16), this interaction may also play an important role in substrate recognition.There is increasing evidence that catalytic domain structures on prothrombin provide an exosite docking surface for prothrombinase (3,17,18). A series of observations suggest that the proexosite I region, the precursor site to IIa exosite I (19), contributes either directly or indirectly to this substrate binding site. For example, reagents that block proexosite I such as hirugen and bothrojaracin inhibit macromolecular substrate cleavage by prothrombinase or the FXa-FVa complex (3, 20, 21). Additionally, proexosite I prethrombin-1 mutants and a naturally occurring prothrombin variant (Arg 67 to His, chymotrypsin numbering system (22)) are poor substrates for prothrombinase and FXa-FVa (23, 24). Interestingly, these probes or mutations reduced the rate of IIa generation only in the presence of the cofactor, suggesting a link between FVa and proexosite I. Consistent with this, IIa exosite I mutants and exosite I probes inhibit FV activation (25-27). Additionally, detailed studies by Bock and co-workers have provided compelling evidence that the FVa heavy chain binds to exosite I of IIa (26,28). Taken together, these data are consistent with the idea that proexosite I plays an important role in macromolecular substrate recogniti...

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“…By contrast, this relationship is much more elusive in dysprothrombinemia and could be related to the molecular defect. 1 We report an homozygous mutation in the prothrombin gene (Arg388(73)His, human prothrombin numbering with the chymotrypsinogen numbering of thrombin residues in brackets) and describe its phenotypic expression.…”

Section: A Natural Variant With a Point Mutation Resulting In A Homozmentioning

confidence: 99%

“…8,9 In conclusion, the experimental data showed that both procoagulant and anticoagulant functions of the R388(73)H natural variant are impaired. Akhavan et al 1 have already proposed that substitutions that affect both procoagulant and anticoagulant functions of thrombin might, at least in part, counterbalance each other so that the hemostatic equilibrium is not drastically modified. The very mild bleeding tendency observed in the proband described here gives further support to this hypothesis.…”

Section: A Natural Variant With a Point Mutation Resulting In A Homozmentioning

confidence: 99%

A natural variant with a point mutation resulting in a homozygous Arg to His substitution at position 388 in prothrombin

d’Audigier¹,

Pasmant²,

Bournier³

et al. 2008

Haematologica

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Molecular and functional characterization of a natural homozygous Arg67His mutation in the prothrombin gene of a patient with a severe procoagulant defect contrasting with a mild hemorrhagic phenotype

Abstract: In a patient who presented with a severe coagulation deficiency in plasma contrasting with a very mild hemorrhagic diathesis a homozygous Arg67His mutation was identified in the prothrombin gene.

Cited by 29 publications

References 37 publications

Role of Prothrombin Fragment 1 in the Pathway of Regulatory Exosite I Formation during Conversion of Human Prothrombin to Thrombin

Role of Prothrombin Fragment 1 in the Pathway of Regulatory Exosite I Formation during Conversion of Human Prothrombin to Thrombin

Role of Hirudin-like Factor Va Heavy Chain Sequences in Prothrombinase Function

A natural variant with a point mutation resulting in a homozygous Arg to His substitution at position 388 in prothrombin

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