Fibrinogen St. Gallen I (γ 292 Gly → Val): Evidence for Structural Alterations Causing Defective Polymerization and Fibrinogenolysis

Stucki, B; Schmutz, Peter; Schmid, L; Haeberli, André; Lämmle, Bernhard; Furlan, Miha

doi:10.1055/s-0037-1614456

Cited by 6 publications

(5 citation statements)

References 16 publications

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“…Interestingly, Stucki et al (57) reported that a mutation that changes Gly292 in human fibrinogen to Val, which is observed in the Fibrinogen St. Gallen I and the Fibrinogen variant Baltimore I, led to an opening of a loop in the backbone chain compared to the structure of normal fibrinogen and the loss of a hydrogen bond, resulting in defective polymerization and fibrinogenolysis. However, this amino acid change is in the middle of an extended turn rather than between an R-helix and a β-strand.…”

Section: Discussionmentioning

confidence: 99%

Unfolding, Aggregation, and Amyloid Formation by the Tetramerization Domain from Mutant p53 Associated with Lung Cancer

et al. 2006

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The p53 tumor suppressor is a tetrameric transcriptional enhancer, and its activity is compromised by mutations that cause amino acid substitutions in its tetramerization domain. Here we analyze the biochemical and biophysical properties of peptides corresponding to amino acids 319 to 358 of wildtype human p53, which includes the tetramerization domain, and that of a cancer-derived mutant with valine substituted for glycine 334. Unlike the wild-type peptide, the G334V peptide forms amyloid fibrils by a two step process under physiological conditions of temperature and pH. Nevertheless, the G334V peptide is capable of forming hetero-oligomers with a wild-type peptide. Computational modeling of the G334V peptide structure suggests that substitution of valine for glycine 334 causes a local distortion that contributes to a β-dominated structural transition leading to amyloid formation. Since the distortion is mostly on the surface, the mutant peptide is still able to form a pseudo-native tetramer complex at higher concentrations and/or lower temperatures. Our † KS was supported in

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Section: Discussionmentioning

confidence: 99%

Unfolding, Aggregation, and Amyloid Formation by the Tetramerization Domain from Mutant p53 Associated with Lung Cancer

et al. 2006

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“…by guest www.bloodjournal.org From both conditions, the digests of fibrinogen Milano XII showed an additional band of apparently higher molecular weight, denoted D1* and D3* in Figure 4A. We compared the plasmin digestion of fibrinogen Milano XII to the digestion of fibrinogen St Gallen I (␥ G292V), 21 a variant that is abnormally digested to give the fragments D2 and D3 because of only partial protection of D1 by Ca ϩϩ . As shown in Figure 4A, fragment D3* was not the same as fragment D2 and thus did not arise from the incomplete conversion of D1 in D3.…”

Section: Sds-page and Immunoblot Analysis Of Fibrinogen Degradation Pmentioning

confidence: 99%

Fibrinogen Milano XII: a dysfunctional variant containing 2 amino acid substitutions, Aα R16C and γ G165R

Bolliger-Stucki

Lord

Furlan

2001

Blood

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Fibrinogen Milano XII was detected in an asymptomatic Italian woman, whose routine coagulation test results revealed a prolonged thrombin time. Fibrinogen levels in functional assays were considerably lower than levels in immunologic assays. Polymerization of purified fibrinogen was strongly impaired in the presence of calcium or ethylenediaminetetraacetic acid (EDTA). Two heterozygous structural defects were detected by DNA analysis: A␣ R16C and ␥ G165R. As seen previously with other heterozygous A␣ R16C variants, thrombin-catalyzed release of fibrinopeptide A was 50% of normal. Additionally, the release of fibrinopeptide B was delayed. Immunoblotting analysis with antibodies to human serum albumin indicated that albumin is bound to A␣ 16 C. Sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) analysis of plasmin digests of fibrinogen Milano XII in the presence of calcium or EDTA showed both normal and novel D1 and D3 fragments. Further digestion of abnormal D3 fragments by chymotrypsin resulted in degradation products of the same size as the fragments derived from normal fibrinogen. SDS-PAGE analysis under reducing conditions showed no difference between normal fibrinogen and fibrinogen Milano XII or between their plasmic fragments. Circular dichroism analysis revealed a shift in the mean residual ellipticity and a significant reduction of the ␣-helix content in the variant D3 fragment. It is concluded that the A␣-chain substitution is mainly responsible for the coagulation abnormalities, whereas the substitution in the ␥-chain induced a conformational change in the D3 fragment. IntroductionFibrinogen, a soluble plasma glycoprotein of 340 kd, is made up of 2 copies of 3 different polypeptide chains (A␣ 2 , B␤ 2 ,␥ 2 ), linked together with 29 interchain and intrachain disulfide bridges. The molecule is organized in a dimeric fashion consisting of a central E domain containing the amino termini of all 6 polypeptide chains and 2 outer D domains. In the final stage of blood coagulation, thrombin cleaves the fibrinopeptides A and B in a sequential manner from the amino termini of the A␣-and B␤-chains. Cleavage occurs between residues R16 (single-letter amino acid abbreviations) and G17 of the A␣-chain and residues R14 and G15 of the B␤-chain, exposing the A and B polymerization sites. Resultant monomers join together to form 2-stranded, halfstaggered protofibrils. It has been shown that protofibrils result from longitudinal D-D interactions and noncovalent contacts between the A and the a sites. The a site is formed by residues 329, 330, 340, and 364 in the carboxy-terminal part of the ␥-chain. 1,2 Subsequently, the growing fibrils aggregate in a lateral fashion to form fibers that increase progressively in thickness and develop branch points. The evolving network is finally stabilized with covalent bonds by the activated factor XIII, resulting in a clot resistant to mechanical disruption.Dysfibrinogenemia is a heritable disorder characterized by structural mutations in any of the 3 polypeptide cha...

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“…In addition, because the novel heterozygous mutation, γA289D, was recently reported as dysfibrinogenemia, the γA289 residue is important for fibrinogen function. The heterozygous γG292V fibrinogens, Baltimore I and St. Gallen I, have been reported as dysfibrinogenemia and hypodysfibrinogenemia, respectively . Because our results indicated that the M/C ratio of recombinant γG292V fibrinogen was lower than that of recombinant WT fibrinogen, we speculated that the γG292V patient manifested hypodysfibrinogenemia with the reduced secretion of variant fibrinogen.…”

Section: Discussionmentioning

confidence: 64%

“…Gallen I, have been reported as dysfibrinogenemia and hypodysfibrinogenemia, respectively. 20,21 Because our results indicated that the M/C ratio of recombinant γG292V fibrinogen was lower than that of recombinant WT fibrinogen, we speculated that the γG292V patient manifested hypodysfibrinogenemia with the reduced secretion of variant fibrinogen. In conclusion, three out of six recombinant variant fibrinogens between γG287R and γG292V in the γ-module showed markedly reduced secretion.…”

Section: Discussionmentioning

confidence: 89%

Heterozygous variant fibrinogen γA289V (Kanazawa III) was confirmed as hypodysfibrinogenemia by plasma and recombinant fibrinogens

Kaido

Yoda

Kamijo

et al. 2020

Int J Lab Hematology

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Introduction Congenital fibrinogen disorders are classified as afibrinogenemia, hypofibrinogenemia, dysfibrinogenemia, and hypodysfibrinogenemia. However, difficulties are associated with discriminating between dysfibrinogenemia, hypofibrinogenemia, and hypodysfibrinogenemia using routine analyses. We previously reported a heterozygous variant fibrinogen (γA289V; Kanazawa III) as hypodysfibrinogenemia; however, the same variant had previously been described as hypofibrinogenemia. To clarify the production of γA289V fibrinogen, we expressed recombinant γA289V (r‐γA289V) fibrinogen and compared it with wild‐type (WT) and adjacent recombinant variant fibrinogens. Methods Target mutations were introduced into a fibrinogen γ‐chain expression vector by site‐directed mutagenesis, and the vector was then transfected into Chinese hamster ovary cells to produce recombinant fibrinogen. Fibrinogen was purified from the plasma of the proposita, and culture media and fibrinogen functions were analyzed using fibrin polymerization, plasmin protection, and FXIIIa‐catalyzed fibrinogen cross‐linking. Results The fibrinogen concentration ratio of the culture media to cell lysates was markedly lower for r‐γA289V fibrinogen than for WT. Because the secretion of recombinant γF290L (r‐γF290L) fibrinogen was similar to WT, we compared r‐γF290L fibrinogen functions with WT. The fibrin polymerization of Kanazawa III plasma (K‐III) fibrinogen was significantly weaker than normal plasma fibrinogen. Moreover, K‐III fibrinogen showed a markedly reduced “D:D” interaction. However, all functions of r‐γF290L fibrinogen were similar to WT. An in silico analysis confirmed the above results. Conclusion The present results demonstrated that γA289 is crucial for the γ‐module structure, and the γA289V substitution markedly reduced fibrinogen secretion. Moreover, K‐III fibrinogen showed markedly reduced fibrin polymerization and “D:D” interactions. γA289V fibrinogen was confirmed as hypodysfibrinogenemia.

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Fibrinogen St. Gallen I (γ 292 Gly → Val): Evidence for Structural Alterations Causing Defective Polymerization and Fibrinogenolysis

Cited by 6 publications

References 16 publications

Unfolding, Aggregation, and Amyloid Formation by the Tetramerization Domain from Mutant p53 Associated with Lung Cancer

Unfolding, Aggregation, and Amyloid Formation by the Tetramerization Domain from Mutant p53 Associated with Lung Cancer

Fibrinogen Milano XII: a dysfunctional variant containing 2 amino acid substitutions, Aα R16C and γ G165R

Heterozygous variant fibrinogen γA289V (Kanazawa III) was confirmed as hypodysfibrinogenemia by plasma and recombinant fibrinogens

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