1. Cell Biology
  2. Structural Biology and Molecular Biophysics
Download icon

Fibrinogen αC-subregions critically contribute blood clot fibre growth, mechanical stability and resistance to fibrinolysis

  1. Helen McPherson
  2. Cedric Duval
  3. Stephen R Baker
  4. Matthew S Hindle
  5. Lih T Cheah
  6. Nathan L Asquith
  7. Marco M Domingues
  8. Victoria C Ridger
  9. Simon DA Connell
  10. Khalid Naseem
  11. Helen Philippou
  12. Ramzi A Ajjan
  13. Robert Ariens  Is a corresponding author
  1. University of Leeds, United Kingdom
  2. Wake Forest University, United States
  3. Harvard Medical School, United States
  4. Universidade de Lisboa, Portugal
  5. University of Sheffield, United Kingdom
  6. University of Leeds, United States
Research Article
  • Cited 0
  • Views 259
  • Annotations
Cite this article as: eLife 2021;10:e68761 doi: 10.7554/eLife.68761

Abstract

Fibrinogen is essential for blood coagulation. The C-terminus of the fibrinogen α-chain (αC-region) is composed of an αC-domain and αC-connector. Two recombinant fibrinogen variants (α390 and α220) were produced to investigate the role of subregions in modulating clot stability and resistance to lysis. The α390 variant, truncated before the αC-domain, produced clots with a denser structure and thinner fibres. In contrast, the α220 variant, truncated at the start of the αC-connector, produced clots that were porous with short, stunted fibres and visible fibre ends. These clots were mechanically weak and susceptible to lysis. Our data demonstrate differential effects for the αC-subregions in fibrin polymerisation, clot mechanical strength, and fibrinolytic susceptibility. Furthermore, we demonstrate that the αC-subregions are key for promoting longitudinal fibre growth. Together, these findings highlight critical functions of the αC-subregions in relation to clot structure and stability, with future implications for development of novel therapeutics for thrombosis.

Data availability

The source data for Figures 1 B-F, figure 2 B and D, figure 3 B, figure 4, figure 5 B, C and D and figure 6 A-C and D-F and supplementary Figures 1 supplement 1, figures 4 supplement 1 and figures 5 supplement 1 and 2 and figures 6 supplement 1 are made available as separate source data files.

Article and author information

Author details

  1. Helen McPherson

    Discovery and Translational Science Department, Leeds Institute of Cariovasular and Metabolic Medicine, University of Leeds, Leeds, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3519-498X
  2. Cedric Duval

    Discovery and Translational Science Department, Leeds Institute of Cariovasular and Metabolic Medicine, University of Leeds, Leeds, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Stephen R Baker

    Department of Physics, Wake Forest University, Winston Salem, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3147-4925
  4. Matthew S Hindle

    Discovery and Translational Science Department, Leeds Institute of Cariovasular and Metabolic Medicine, University of Leeds, Leeds, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  5. Lih T Cheah

    Discovery and Translational Science Department, Leeds Institute of Cariovasular and Metabolic Medicine, University of Leeds, Leeds, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  6. Nathan L Asquith

    Division of Hematology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Marco M Domingues

    Instituto de Medicina Molecular - João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, Lisbon, Portugal
    Competing interests
    The authors declare that no competing interests exist.
  8. Victoria C Ridger

    Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  9. Simon DA Connell

    Molecular and Nanoscale Physics Group, University of Leeds, Leeds, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Khalid Naseem

    Discovery and Translational Science Department, University of Leeds, Leeds, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  11. Helen Philippou

    Discovery and Translational Science Department, Leeds Institute of Cariovasular and Metabolic Medicine, University of Leeds, Leeds, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  12. Ramzi A Ajjan

    Discovery and Translational Science Department, Leeds Institute of Cariovasular and Metabolic Medicine, University of Leeds, Leeds, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1636-3725
  13. Robert Ariens

    Discovery anTranslational Science Department, University of Leeds, Leeds, United Kingdom
    For correspondence
    R.A.S.Ariens@leeds.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6310-5745

Funding

British Heart Foundation (RG/13/3/30104)

  • Helen McPherson
  • Cedric Duval
  • Stephen R Baker
  • Marco M Domingues
  • Victoria C Ridger
  • Simon DA Connell
  • Helen Philippou
  • Ramzi A Ajjan
  • Robert Ariens

British Heart Foundation (RG/18/11/34036)

  • Helen McPherson
  • Cedric Duval
  • Stephen R Baker
  • Victoria C Ridger
  • Simon DA Connell
  • Helen Philippou
  • Ramzi A Ajjan
  • Robert Ariens

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: Procedures were performed according to accepted standards of humane animal care, approved by the ethical review committee at the University of Leeds, and conducted under license (P144DD0D6) from the United Kingdom Home Office.

Reviewing Editor

  1. Jameel Iqbal, Icahn School of Medicine at Mount Sinai, United States

Publication history

  1. Received: March 24, 2021
  2. Preprint posted: May 8, 2021 (view preprint)
  3. Accepted: October 4, 2021
  4. Accepted Manuscript published: October 11, 2021 (version 1)
  5. Accepted Manuscript updated: October 15, 2021 (version 2)
  6. Version of Record published: October 28, 2021 (version 3)

Copyright

© 2021, McPherson et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 259
    Page views
  • 61
    Downloads
  • 0
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Cell Biology
    Julie Favre et al.
    Research Article

    Estrogen receptor alpha (ERα) activation by estrogens prevents atheroma through its nuclear action whereas plasma membrane-located ERα accelerates endothelial healing. The genetic deficiency of ERα was associated with a reduction in flow-mediated dilation (FMD) in one man. Here, we evaluated ex vivo the role of ERα on FMD of resistance arteries. FMD, but not agonist (acetylcholine, insulin)-mediated dilation, was reduced in male and female mice lacking ERα (Esr1-/- mice) compared to wild-type mice and was not dependent on the presence of estrogens. In C451A-ERα mice lacking membrane ERα, not in mice lacking AF2-dependent nuclear ERα actions, FMD was reduced, and restored by antioxidant treatments. Compared to wild-type mice, isolated perfused kidneys of C451A-ERα mice revealed a decreased flow-mediated nitrate production and an increased H2O2 production. Thus, endothelial membrane ERα promotes NO bioavailability through inhibition of oxidative stress and thereby participates in FMD in a ligand-independent manner.

    1. Biochemistry and Chemical Biology
    2. Cell Biology
    Rania Elsabrouty et al.
    Research Article

    UbiA prenyltransferase domain-containing protein-1 (UBIAD1) utilizes geranylgeranyl pyrophosphate (GGpp) to synthesize the vitamin K2 subtype menaquinone-4. The prenyltransferase has emerged as a key regulator of sterol-accelerated, endoplasmic reticulum (ER)-associated degradation (ERAD) of HMG CoA reductase, the rate-limiting enzyme in synthesis of cholesterol and nonsterol isoprenoids including GGpp. Sterols induce binding of UBIAD1 to reductase, inhibiting its ERAD. Geranylgeraniol (GGOH), the alcohol derivative of GGpp, disrupts this binding and thereby stimulates ERAD of reductase and translocation of UBIAD1 to Golgi. We now show that overexpression of Type 1 polyisoprenoid diphosphate phosphatase (PDP1), which dephosphorylates GGpp and other isoprenyl pyrophosphates to corresponding isoprenols, abolishes protein geranylgeranylation as well as GGOH-induced ERAD of reductase and Golgi transport of UBIAD1. Conversely, these reactions are enhanced in the absence of PDP1. Our findings indicate PDP1-mediated hydrolysis of GGpp significantly contributes to a feedback mechanism that maintains optimal intracellular levels of the nonsterol isoprenoid.