Automated, Microscopic Measurement of Fibrinaloid Microclots and Their Degradation by Nattokinase, the Main Natto Protease

Authors

  • Dr. Justine M. Grixti Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L69 7ZB, UK Author
  • Dr. Chrispian W. Theron Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L69 7ZB, UK; Gene Mill Biofoundry, Institute of Systems, Molecular and Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L69 7ZB, UK Author
  • Dr. J. Enrique Salcedo-Sora Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L69 7ZB, UK; Gene Mill Biofoundry, Institute of Systems, Molecular and Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L69 7ZB, UK Author
  • Prof. Etheresia Pretorius Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L69 7ZB, UK; Department of Physiological Sciences, Faculty of Science, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa Author https://orcid.org/0000-0002-9108-2384
  • Douglas B. Kell Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L69 7ZB, UK; Gene Mill Biofoundry, Institute of Systems, Molecular and Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L69 7ZB, UK; Department of Physiological Sciences, Faculty of Science, Stellenbosch University, Private Bag X1, Matieland 7602, South Africa; The Novo Nordisk Foundation Centre for Biosustainability, Technical University of Denmark, Søltofts Plads 220, 2800 Kgs Lyngby, Denmark Author

DOI:

https://doi.org/10.62767/jecacm504.6557

Keywords:

microclots, fibrinaloid, chronic disease, fibrinolysis, nattokinase

Abstract

Nattokinase, from the Japanese fermented food natto, is a protease with fibrinolytic activity that can thus degrade conventional blood clots. In some cases, however, including in Long COVID, fibrinogen can polymerise into an anomalous amyloid form to create clots that are resistant to normal fibrinolysis and that we refer to as fibrinaloid microclots. These can be detected with the fluorogenic stain thioflavin T. We describe an automated microscopic technique for the quantification of fibrinaloid microclot formation, which also allows the kinetics of their formation and aggregation to be recorded. We also here show that recombinant nattokinase is effective at degrading the fibrinaloid microclots in vitro. Flow conditions, mimicked by shaking, increase the size of the clots via aggregation. Overall, this work adds to the otherwise largely anecdotal evidence, that we review, that nattokinase might be anticipated to have value as part of therapeutic treatments for individuals with Long COVID and related disorders that involve fibrinaloid microclots.

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Published

2024-11-21

Data Availability Statement

All data are available either in Supplementary Information or on request from the first author.

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