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Comparison of molecular structure and fibrin polymerization between two Bβ-chain N-terminal region fibrinogen variants, Bβp.G45C and Bβp.R74C

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Abstract

We identified two heterozygous dysfibrinogenemias, Bβp.Gly45Cys (Kyoto VII; K-VII) and Bβp.Arg74Cys (Iida II; I-II). The impairment of polymerization of Bβp.G45C has been well analyzed; however, that of Bβp.R74C has not. Thus, we compared fibrin polymerization between these variants. To determine the structural and functional characterization of purified fibrinogens, we performed immunoblotting analysis, kinetic analyses of fibrinopeptide A and B release, and thrombin- or batroxobin-catalyzed fibrin or fibrin monomer polymerization. Immunoblotting analysis showed that both variant fibrinogens had variant fibrinogen-albumin complexes and variant fibrinogen multimers, and the amounts of fibrinogen-albumin complexes with fibrinogen K-VII was more than with fibrinogen I-II. Moreover, fibrinopeptide B release from fibrinogen K-VII was about 50% of the control, whereas the others were normal. The maximum slopes of polymerization for variant fibrinogens were reduced, but fibrinogen K-VII was reduced more than fibrinogen I-II. The present study demonstrated that both Bβp.G45C and Bβp.R74C variants showed the presence of variant fibrinogen-albumin complexes and variant fibrinogen multimers, and polymerization of Bβp.G45C was impaired more than Bβp.R74C. Our study and several previous reports concerning the clinical phenotype of both variants suggested the risks of bleeding for patients with Bβp.G45C and thrombosis for patients with Bβp.R74C.

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Acknowledgements

We gratefully acknowledge Dr. Toru Inaba (Kyoto Prefectural University of Medicine, Kyoto) and Dr. Yu Furui (Shinshu University of School of Medicine, Matsumoto) for the patient referrals. This work was supported by JSPS KAKENHI Grant Number JP17K09009 (Chiaki Taira and Nobuo Okumura).

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Authors and Affiliations

  1. Department of Clinical Laboratory Investigation, Graduate School of Medicine, Shinshu University, Matsumoto, Japan

    Takahiro Kaido, Masahiro Yoda, Tomu Kamijo, Chiaki Taira, Yumiko Higuchi & Nobuo Okumura

  2. Laboratory of Clinical Chemistry and Immunology, Department of Biomedical Laboratory Sciences, School of Health Sciences, Shinshu University, 3-1-1 Asahi, Matsumoto, 390-8621, Japan

    Nobuo Okumura

Authors
  1. Takahiro Kaido
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  2. Masahiro Yoda
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  3. Tomu Kamijo
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  4. Chiaki Taira
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  5. Yumiko Higuchi
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  6. Nobuo Okumura
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Contributions

TK and MY performed the research, analyzed the data. TK wrote the manuscript. TK, CT, YH, and NO designed the research and discussed the data. NO and TK reviewed the manuscript.

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Correspondence to Nobuo Okumura.

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12185_2020_2919_MOESM1_ESM.tif

Supplementary file1 (TIF 1194 kb) Supplemental Figure 1: Western blotting analysis under reducing conditions using an anti-human albumin antibody and an anti-human fibrinogen antibody. Purified plasma fibrinogens were analyzed by western blotting in reducing conditions (Panel A: anti-human albumin antibody, Panel B: anti-human fibrinogen antibody). Panel A: the band that reacted with anti-human albumin antibody was shown in both variant fibrinogens. Panel B: these bands showed the typical pattern. Above the dashed line: stacking gel, below the dashed line: resolving gel. 1: normal fibrinogen, 2: fibrinogen Kyoto VII (Bβp.G45C), 3: fibrinogen Iida II (Bβp.R74C). Alb: albumin.

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Kaido, T., Yoda, M., Kamijo, T. et al. Comparison of molecular structure and fibrin polymerization between two Bβ-chain N-terminal region fibrinogen variants, Bβp.G45C and Bβp.R74C. Int J Hematol 112, 331–340 (2020). https://doi.org/10.1007/s12185-020-02919-5

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  • DOI: https://doi.org/10.1007/s12185-020-02919-5

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