Calcium‐binding properties of human fibrin(ogen) and degradation products

Nieuwenhuizen, W.; Vermond, Anton; Nooijen, W. J.; Haverkate, F.

doi:10.1016/0014-5793(79)80194-5

Cited by 43 publications

(14 citation statements)

References 9 publications

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“…On the basis of their finding that the thrombin-fibrin interaction is sensitive to the presence of Ca2+ ions and that thrombin is released at some stage of fibrin polymerization, Kaminski and McDonagh (1983) speculated that it might be the D domain of fibrinogen that carries an extended thrombin binding site, since this domain contains both polymerization sites (Olexa & Budzynski, 1981; Varadi & Scheraga, 1986) and Ca2+ binding sites (Nieuwenhuizen et al, 1979;Varadi & Scheraga, 1986). However, at present it is not known when thrombin is released from fibrin (see later section).…”

Section: Discussionmentioning

confidence: 99%

Localization of the binding site on fibrin for the secondary binding site of thrombin

Vali

Scheraga²

1988

Biochemistry

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Affinity chromatography of active site inhibited thrombin on immobilized fragments derived from the central (desAB-NDSK) and terminal (D1) globular domains of fibrinogen revealed that the site responsible for the binding of thrombin at its secondary fibrin binding site is located in the central domain. Chromatography of various domains of the central nodule (desAB-NDSK, fibrinogen E, and fibrin E) having nonidentical amino acid sequences showed that all of these fragments are capable of binding to PMSF-thrombin-Sepharose, suggesting that the thrombin binding site resides within the peptide regions common to all of these fragments: alpha(Gly17-Met51), beta(Val55-Met118), and gamma(Tyr1-Lys53). Competitive affinity chromatography of the same binding domains revealed that there is no detectable difference in their binding constants to PMSF-thrombin-Sepharose, indicating that the alpha(Lys52-Lys78), beta(Gly15-Lys54)/(Tyr119-Lys122), and gamma(Thr54-Met78) peptide segments do not contribute significantly to the binding of thrombin. Chromatography of the isolated chains of fibrinogen E showed that the alpha(Gly17-Lys78) peptide region itself contains a strong binding site for PMSF-thrombin-Sepharose. The location of the binding site suggests that the secondary site interaction may play an important role in determining the cleavage specificity of thrombin on fibrinogen and can affect the rate of release of the fibrinopeptides. Affinity chromatography of fragments prepared from polymerized fibrin showed that cross-linked DD (D x D) itself does not bind to thrombin, whereas the D x DE complex remained attached to the column, suggesting that the binding site on fragment E for thrombin is distinct from its binding site for D x D.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Discussionmentioning

confidence: 99%

Localization of the binding site on fibrin for the secondary binding site of thrombin

Vali

Scheraga²

1988

Biochemistry

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“…There may be as many as three high-affinity cal cium binding sites per molecule of fibrinogen and 10-12 low-affinity sites [50]. One high-affinity site is located in each of the D-domains [51], and has K d = 9 x 10 -6 M, while K d of the reported third site is 5 times larger. Binding of calcium to these highaffinity sites can obviously not be related to regula tory function.…”

Section: Interactions Of Fibrinogen-fibrin With Other Moleculesmentioning

confidence: 99%

Fibrin: Structure and Interactions

Hermans¹,

McDonagh²

1982

Semin Thromb Hemost

123

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“…It has been suggested (4) that the carboxy termini of the Aa-chains might be responsible for binding of the third Ca2+ ion. Nieuwenhuizen et al (5) ascribed the third Ca2+ binding site to the amino terminal disulfide knot of the fibrinogen molecule although there was no calcium binding noted in fragment E (2,6). While the low affin ity binding sites have been tentatively attributed to sialic acid residues (7-9) situated in the carbohydrate side-chains of the B(3-and 7 -chains, a peptide from the 7 -chain of human fibrinogen capable of binding Ca2+ with high affinity was localized in the D-domain between positions 7 311-336 (10).…”

Section: Introductionmentioning

confidence: 99%

Normal Binding of Calcium to Five Fibrinogen Variants with Mutations in the Carboxy Terminal Part of the γ-Chain

et al. 1996

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SummaryCalcium ions are known to accelerate polymerization of fibrin monomers. Each of the two carboxy terminal domains of normal fibrinogen contains one high-affinity calcium binding site that seems to be situated close to the polymerization site in the γ-chain. Most hitherto described functionally defective fibrinogen variants showed impaired clot formation. Since the tightly bound calcium ions may influence the conformation of the polymerization site, the question arises whether the abnormal clotting of a dysfibrinogen might be due to defective calcium binding. We investigated binding of calcium to fibrinogen and the effect of calcium on the clotting properties of five heterozygous fibrinogen variants showing normal thrombin-induced fibrinopeptide release but abnormal polymerization of fibrin monomers. Each of these dysfibrinogens has one single amino acid substitution in the carboxy-terminal part of the γ-chain: fibrinogen Claro (γ 275 Arg ⟶ His), Milano V (γ 275 Arg Cys), Milano I (γ 330 Asp ⟶ Val), Bern I (γ 337 Asn Lys), and Milano VII (γ 358 Ser Cys). The shortest thrombin clotting time and the earliest onset of turbidity increase were observed in fibrinogen γ 358 Ser ⟶ Cys; both parameters were little affected by calcium concentration. In the variant γ 337 Asn ⟶ Lys, the thrombin time was abnormally prolonged at 0.01 mM Ca2+, but it was normalized at 1 mM calcium. In contrast, the abnormal fibrin polymerization of fibrinogen γ 330 Asp ⟶ Val was barely improved at increasing calcium concentrations. Both variants with the substitution of γ 275 Arg, the residue indispensable for normal D:D interactions, showed the slowest rate of fibrin polymerization and the lowest turbidity of fibrin clots at any Ca2+ concentration used. High affinity calcium binding was found to be normal in all five fibrinogen variants studied, suggesting that their abnormal clotting was not due to defective binding of calcium. The γ-chain in the fragment D1 derived from the variant γ 337 Asn ⟶ Lys was further degraded by plasmin in the presence and in the absence of calcium, whereas fragments D1 from the other four γ-chain variants as well as from normal fibrinogen were protected against plasmic degradation in the presence of 1 mM Ca2+.

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Calcium‐binding properties of human fibrin(ogen) and degradation products

Cited by 43 publications

References 9 publications

Localization of the binding site on fibrin for the secondary binding site of thrombin

Localization of the binding site on fibrin for the secondary binding site of thrombin

Fibrin: Structure and Interactions

Normal Binding of Calcium to Five Fibrinogen Variants with Mutations in the Carboxy Terminal Part of the γ-Chain

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