Important role of arginine 129 in heparin-binding site of antithrombin III. Identification of a novel mutation arginine 129 to glutamine.

Gandrille, Sophie; Aiach, Martine; Lane, David A.; Vidaud, Dominique; Molho-Sabatier, P; Caso, R; Moerloose, P de; Fiessinger, Jean‐Noël; Clauser, Éric

doi:10.1016/s0021-9258(17)30614-2

Cited by 54 publications

(11 citation statements)

References 38 publications

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“…Recently, we have investigated the properties of variants of human antithrombin with abnormal heparin affinity (Owen et al, 1987;Brennan et al, 1988;Gandrille et al, 1990) and have also studied the heparin binding properties of novel conformational forms of antithrombin (Carrell et al, 1991).…”

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confidence: 99%

Heparin binding site, conformational change, and activation of antithrombin

Evans¹,

Marshall²,

Christey³

et al. 1992

Biochemistry

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Alignment of the heparin-activated serpins indicates the presence of two binding sites for heparin: a small high-affinity site on the D-helix corresponding in size to the minimal pentasaccharide heparin, and a longer contiguous low-affinity site extending to the reactive center pole of the molecule. Studies of the complexing of antithrombin and its variants with heparin fractions and with reactive center loop peptides including intermolecular loop-sheet polymers all support a 3-fold mechanism for the heparin activation of antithrombin. Binding to the pentasaccharide site induces a conformational change as measured by circular dichroism. Accompanying this, the reactive center becomes more accessible to proteolytic cleavage and there is a 100-fold increase in the kass for factor Xa but only a 10-fold increase for thrombin, to 6.4 x 10(4) M-1 s-1. To obtain a 100-fold increase in the kass for thrombin requires in addition a 4:1 molar ratio of disaccharide to neutralize the charge on the extended low-affinity site. Full activation requires longer heparin chains in order to stabilize the ternary complex between antithrombin and thrombin. Thus, addition of low-affinity but high molecular weight heparin in conjunction with pentasaccharide gives an overall kass of 2.7 x 10(6) M-1 s-1, close to that of maximal heparin activation.

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confidence: 99%

Heparin binding site, conformational change, and activation of antithrombin

Evans¹,

Marshall²,

Christey³

et al. 1992

Biochemistry

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“…Evidence for the involvement in heparin binding of residues in these helices has come from chemical modification experiments (Blackburn et al, 1984;Chang, 1989;Sun & Chang, 1990) and from studying plasma from patients with known mutations. Studies have been reported on natural substitution mutations of amino acids in the heparin binding domain: the structurally important Pro41 in AT III Basel (Chang & Tran, 1986) and the positively charged Arg47 in AT III Padua 2 (Olds et al, 1990), Alger (Wolf et al, 1982), Paris (Fischer et al, 1986), and Rouen I (Owen et al, 1987) as well as Arg 129 in AT III Geneva (Gandrille et al, 1990). In one of these studies, the variant AT III Geneva (Arg 129 to Gin) was separated from plasma by its nonbinding to heparin-Sepharose in 0.4 M NaCl but was shown to have some residual heparin cofactor activity.…”

Section: Discussionmentioning

confidence: 99%

“…Heparin-Sepharose chromatography, as described previously, was also used to separate variants AT III Geneva (Gandrille et al, 1990), Budapest 1 (Olds et al, 1992b), and Budapest 5 (Lane et al, 1992) from patient plasmas.…”

Section: Methodsmentioning

confidence: 99%

Heparin binding affinity of normal and genetically modified antithrombin III measured using a monoclonal antibody to the heparin binding site of antithrombin III

Watton

Longstaff²,

Lane³

et al. 1993

Biochemistry

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The inhibitory activity of the plasma serine proteinase inhibitor antithrombin III (AT III) is enhanced about 1000-fold upon binding to heparin. We have determined the dissociation constants, Kd, of 48.8 nM for the heparin-AT III interaction, of 175 nM for the specific pentasaccharide-AT III interaction, and of 13 microM for the low-affinity heparin-AT III interaction, using a binding assay based on a monoclonal antibody (MAb) that recognizes an epitope at or close to the heparin binding site of AT III. The heparin binding affinities and proportions of normal and variant AT III in plasma from patients with mutations of AT III have been quantitated for the first time using the binding assay. Substitution mutations in three regions of AT III have been investigated: (i) mutations in the reactive site loop affecting Ala382, Arg393, and Ser394 have no discernible effect on heparin binding; (ii) mutations in the previously identified N-terminal heparin binding region, affecting Arg47, Leu99, and Arg129, produce variant AT III molecules with heparin affinities reduced 11-924-fold, the largest reduction being observed for the substitution mutation Arg47-Cys in Padua 2, which has an affinity of 65.6 microM; (iii) mutations in the hydrophobic regions around strand 1C of the C terminus, affecting Phe402, Ala404, Asn405, Pro407, and Pro429, have pleiotropic effects that include the production of reduced amounts of low-affinity AT III with dissociation constants from 6 to 43 microM.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…However, SERPINC1 c.1154-14G>A variant is well-demonstrated, inducing type 1 deficiency (15). The missense mutations of amino acid 114 to 156, the region involved in heparin-binding, could lead to type 2 deficiency (16,17). As mentioned above, a variant of p. Arg79Cys causes type 2 deficiency (12).…”

Section: Discussionmentioning

confidence: 98%

Utility of the SERPINC1 Gene Test in Ischemic Stroke Patients With Antithrombin Deficiency

et al. 2022

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ObjectiveAntithrombin (AT) plays a critical role in the coagulation system, and its deficiency induces hypercoagulability. AT deficiency is caused not only by inherited variants in the SERPINC1 gene but also by acquired conditions. Therefore, AT deficiency alone could not ensure the presence of the SERPINC1 mutation. We evaluated the utility of the SERPINC1 gene test in ischemic stroke, an important clinical type of arterial thrombosis.MethodsThis retrospective, observational study investigated symptomatic patients who underwent the SERPINC1 gene test because of decreased AT activity (<80%) during 2009-2021 at a tertiary hospital. For the detection of sequence variants in the SERPINC1 gene, direct Sanger sequencing and multiplex ligation-dependent probe amplification were performed. The phenotypes of patients with SERPINC1 gene mutations were examined, and the conditions associated with the pathogenic variants were analyzed.ResultsIn our cohort (n = 19), 13 of 19 patients (68.4%) had the pathogenic variant of the SERPINC1 gene. Ischemic stroke (n = 7) was significantly associated with the pathogenic variants (p = 0.044), and the pathogenicity detection rate was 100%. For any kind of arterial thrombosis (n = 8), the detection rate of the pathogenic variant was 87.5%, but was not statistically significant (p = 0.177). The detection rates of the pathogenic variant in ischemic stroke or arterial thrombosis groups were both higher than those in the venous thrombosis-only group (54.5%).ConclusionThe SERPINC1 gene test was useful in determining the cause of AT deficiency-related arterial thrombosis, especially ischemic stroke. We propose the diagnostic flow of SERPINC1-related ischemic stroke.

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Important role of arginine 129 in heparin-binding site of antithrombin III. Identification of a novel mutation arginine 129 to glutamine.

Cited by 54 publications

References 38 publications

Heparin binding site, conformational change, and activation of antithrombin

Heparin binding site, conformational change, and activation of antithrombin

Heparin binding affinity of normal and genetically modified antithrombin III measured using a monoclonal antibody to the heparin binding site of antithrombin III

Utility of the SERPINC1 Gene Test in Ischemic Stroke Patients With Antithrombin Deficiency

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