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A novel variant fibrinogen, AαE11del, demonstrating the importance of AαE11 residue in thrombin binding

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Abstract

Introduction

We identified a novel heterozygous AαE11del variant in a patient with congenital dysfibrinogenemia. This mutation is located in fibrinopeptide A (FpA). We analyzed the effect of AαE11del on the catalyzation of thrombin and batroxobin and simulated the stability of the complex structure between the FpA fragment (AαG6-V20) peptide and thrombin.

Materials and methods

We performed fibrin polymerization and examined the kinetics of FpA release catalyzed by thrombin and batroxobin using purified plasma fibrinogen. To clarify the association between the AαE11 residue and thrombin, we calculated binding free energy using molecular dynamics simulation trajectories.

Results

Increasing the thrombin concentration improved release of FpA from the patient’s fibrinogen to approximately 90%, compared to the previous 50% of that of normal fibrinogen. Fibrin polymerization of variant fibrinogen also improved. In addition, greater impairment of variant FpA release from the patient’s fibrinogen was observed with thrombin than with batroxobin. Moreover, the calculated binding free energy showed that the FpA fragment–thrombin complex became unstable due to the missing AαE11 residue.

Conclusions

Our findings indicate that the AαE11 residue is involved in FpA release in thrombin catalyzation more than in batroxobin catalyzation, and that the AαE11 residue stabilizes FpA fragment–thrombin complex formation.

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Acknowledgements

We gratefully acknowledge Dr. Takeshi Sato (Soka Municipal Hospital, Soka, Saitama), for the patient referral. This work was supported by JSPS KAKENHI Grant Number JP20K07799 (Nobuo Okumura).

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Authors and Affiliations

  1. Department of Medical Sciences, Graduate School of Medicine, Science and Technology, Shinshu University, Matsumoto, Japan

    Takahiro Kaido & Tomu Kamijo

  2. Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan

    Takahiro Kaido & Tomu Kamijo

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

    Masahiro Yoda, Shinpei Arai, Kazuyoshi Yamauchi & Nobuo Okumura

  4. 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. Shinpei Arai
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  5. Kazuyoshi Yamauchi
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  6. Nobuo Okumura
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Contributions

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

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Correspondence to Nobuo Okumura.

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Kaido, T., Yoda, M., Kamijo, T. et al. A novel variant fibrinogen, AαE11del, demonstrating the importance of AαE11 residue in thrombin binding. Int J Hematol 114, 591–598 (2021). https://doi.org/10.1007/s12185-021-03200-z

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  • DOI: https://doi.org/10.1007/s12185-021-03200-z

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