Fractal kinetic behavior of plasmin on the surface of fibrin meshwork

Imre Varjú, Kiril Tenekedjiev, Z. Keresztes, A. Pap, László Szabó, Craig Thelwell, Colin Longstaff, R. Machovich, K. Kolev

Research output: Article

10 Citations (Scopus)

Abstract

Intravascular fibrin clots are resolved by plasmin acting at the interface of gel-phase substrate and fluid-borne enzyme. The classic Michaelis-Menten kinetic scheme cannot describe satisfactorily this heterogeneous-phase proteolysis because it assumes homogeneous well-mixed conditions. A more suitable model for these spatial constraints, known as fractal kinetics, includes a time-dependence of the Michaelis coefficient KmF = Km0F(1 + t)h, where h is a fractal exponent of time, t. The aim of the present study was to build up and experimentally validate a mathematical model for surface-acting plasmin that can contribute to a better understanding of the factors that influence fibrinolytic rates. The kinetic model was fitted to turbidimetric data for fibrinolysis under various conditions. The model predicted Km0F= 1.98 μM and h = 0.25 for fibrin composed of thin fibers and Km0F= 5.01 μM and h = 0.16 for thick fibers in line with a slower macroscale lytic rate (due to a stronger clustering trend reflected in the h value) despite faster cleavage of individual thin fibers (seen as lower Km0F). ε-Aminocaproic acid at 1 mM or 8 U/mL carboxypeptidase-B eliminated the time-dependence of KmF and increased the lysis rate suggesting a role of C-terminal lysines in the progressive clustering of plasmin. This fractal kinetic concept gained structural support from imaging techniques. Atomic force microscopy revealed significant changes in plasmin distribution on a patterned fibrinogen surface in line with the time-dependent clustering of fluorescent plasminogen in confocal laser microscopy. These data from complementary approaches support a mechanism for loss of plasmin activity resulting from C-terminal lysine-dependent redistribution of enzyme molecules on the fibrin surface.

Original languageEnglish
Pages (from-to)6348-6356
Number of pages9
JournalBiochemistry
Volume53
Issue number40
DOIs
Publication statusPublished - okt. 14 2014

Fingerprint

Fractals
Fibrinolysin
Fibrin
Kinetics
Cluster Analysis
Confocal Microscopy
Lysine
Fibers
Carboxypeptidase B
Proteolysis
Aminocaproic Acid
Enzyme kinetics
Atomic Force Microscopy
Plasminogen
Fibrinolysis
Enzymes
Fibrinogen
Atomic force microscopy
Microscopic examination
Theoretical Models

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)

Cite this

Fractal kinetic behavior of plasmin on the surface of fibrin meshwork. / Varjú, Imre; Tenekedjiev, Kiril; Keresztes, Z.; Pap, A.; Szabó, László; Thelwell, Craig; Longstaff, Colin; Machovich, R.; Kolev, K.

In: Biochemistry, Vol. 53, No. 40, 14.10.2014, p. 6348-6356.

Research output: Article

Varjú, I, Tenekedjiev, K, Keresztes, Z, Pap, A, Szabó, L, Thelwell, C, Longstaff, C, Machovich, R & Kolev, K 2014, 'Fractal kinetic behavior of plasmin on the surface of fibrin meshwork', Biochemistry, vol. 53, no. 40, pp. 6348-6356. https://doi.org/10.1021/bi500661m
Varjú, Imre ; Tenekedjiev, Kiril ; Keresztes, Z. ; Pap, A. ; Szabó, László ; Thelwell, Craig ; Longstaff, Colin ; Machovich, R. ; Kolev, K. / Fractal kinetic behavior of plasmin on the surface of fibrin meshwork. In: Biochemistry. 2014 ; Vol. 53, No. 40. pp. 6348-6356.
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AU - Szabó, László

AU - Thelwell, Craig

AU - Longstaff, Colin

AU - Machovich, R.

AU - Kolev, K.

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