Free fatty acids modulate thrombin mediated fibrin generation resulting in less stable clots

Anna Tanka-Salamon, Erzsébet Komorowicz, László Szabó, Kiril Tenekedjiev, K. Kolev

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Upon platelet activation, free fatty acids are released at the stage of thrombus formation, but their effects on fibrin formation are largely unexplored. Our objective was to characterize the kinetic effects of fatty acids on thrombin activity, as well as the structural and mechanical properties of the resultant fibrin clots. Thrombin activity on fibrinogen was followed by turbidimetry and detailed kinetic characterization was performed using a fluorogenic short peptide substrate. The viscoelastic properties of fibrin were measured with rotatory oscillation rheometer, whereas its structure was analyzed with scanning electron microscopy (SEM). In turbidimetric assays of fibrin generation, oleate and stearate at physiologically relevant concentrations (60-600 μM) produced a bell-shaped inhibitory dose response, increasing 10- to 30-fold the time to half-maximal clotting. Oleate inhibited thrombin activity on a short peptide substrate according to a mixed-type inhibitor pattern (a 9-fold increase of the Michaelis constant, Km and a 20% decrease of the catalytic constant), whereas stearate resulted in only a minor (15%) drop in the catalytic constant without any change in the Km . Morphometric analysis of SEM images showed a 73% increase in the median fiber diameter in the presence of stearate and a 20% decrease in the presence of oleate. Concerning the viscoelastic parameters of the clots, storage and loss moduli, maximal viscosity and critical shear stress decreased by 32-65% in the presence of oleate or stearate, but loss tangent did not change indicating decreased rigidity, higher deformability and decreased internal resistance to shear stress. Our study provides evidence that free fatty acids (at concentrations comparable to those reported in thrombi) reduce the mechanical stability of fibrin through modulation of thrombin activity and the pattern of fibrin assembly.

Original languageEnglish
Article numbere0167806
JournalPLoS One
Volume11
Issue number12
DOIs
Publication statusPublished - Dec 1 2016

Fingerprint

fibrin
thrombin
Fibrin
Nonesterified Fatty Acids
Thrombin
free fatty acids
Stearates
Oleic Acid
stearic acid
oleic acid
Electron Scanning Microscopy
shear stress
Shear stress
Thrombosis
scanning electron microscopy
peptides
Nephelometry and Turbidimetry
platelet activation
kinetics
loss modulus

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Free fatty acids modulate thrombin mediated fibrin generation resulting in less stable clots. / Tanka-Salamon, Anna; Komorowicz, Erzsébet; Szabó, László; Tenekedjiev, Kiril; Kolev, K.

In: PLoS One, Vol. 11, No. 12, e0167806, 01.12.2016.

Research output: Contribution to journalArticle

Tanka-Salamon, Anna ; Komorowicz, Erzsébet ; Szabó, László ; Tenekedjiev, Kiril ; Kolev, K. / Free fatty acids modulate thrombin mediated fibrin generation resulting in less stable clots. In: PLoS One. 2016 ; Vol. 11, No. 12.
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