Polyphosphate modifies the fibrin network and down-regulates fibrinolysis by attenuating binding of tPA and plasminogen to fibrin

Nicola J Mutch, Ruchira Engel, Shirley Uitte de Willige, Helen Philippou, Robert A S Ariëns

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

Activated platelets secrete a negatively charged polymer, polyphosphate (polyP). Here, we explore the interactions of polyP with fibrin(ogen) and its effect on fibrin structure and fibrinolysis. Electrophoretic mobility and binding assays indicate that polyP interacts with fibrinogen and soluble fibrin. Clots formed in the presence of polyP exhibited reduced turbidity and permeability indicative of a tighter fibrin network, but these changes were not related to cross-linking or fibrinopeptide release. Microscopy showed a change in fibrin distribution in clots formed with polyP; with formation of tight aggregates of fibrin fibers interspaced with large pores in contrast to homogenous fiber distribution in control clots. Lysis by tissue plasminogen activator (tPA) and plasminogen or plasmin was delayed in clots formed with polyP and depended on both the activator and polyP concentration. Adding polyP to the clot after fibrin formation or to repolymerizing soluble fibrin did not affect lysis, indicating changes induced by polyP occur at the level of conversion of fibrinogen to fibrin. Surface plasmon resonance showed that the presence of polyP reduced the binding of both plasminogen and tPA to partially lysed fibrin surfaces. These data show that polyP directly influences fibrin architecture and attenuates fibrinolysis through reduced binding of fibrinolytic proteins.
Original languageEnglish
Pages (from-to)3980-3988
Number of pages9
JournalBlood
Volume115
Issue number19
Early online date12 Mar 2010
DOIs
Publication statusPublished - 13 May 2010

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Polyphosphates
Plasminogen
Fibrinolysis
Tissue Plasminogen Activator
Fibrin
Polymers
Down-Regulation
Fibrinogen
Electrophoretic mobility
Surface Plasmon Resonance
Fibers
Fibrinolysin
Surface plasmon resonance
Turbidity
Platelets
Microscopy
Permeability
Assays
Carrier Proteins
Microscopic examination

Keywords

  • down-regulation
  • electrophoretic mobility shift assay
  • fibrin
  • fibrinolysin
  • fibrinolysis
  • humans
  • plasminogen
  • polyphosphates
  • surface plasmon resonance
  • tissue plasminogen activator

Cite this

Polyphosphate modifies the fibrin network and down-regulates fibrinolysis by attenuating binding of tPA and plasminogen to fibrin. / Mutch, Nicola J; Engel, Ruchira; Uitte de Willige, Shirley; Philippou, Helen; Ariëns, Robert A S.

In: Blood, Vol. 115, No. 19, 13.05.2010, p. 3980-3988.

Research output: Contribution to journalArticle

Mutch, Nicola J ; Engel, Ruchira ; Uitte de Willige, Shirley ; Philippou, Helen ; Ariëns, Robert A S. / Polyphosphate modifies the fibrin network and down-regulates fibrinolysis by attenuating binding of tPA and plasminogen to fibrin. In: Blood. 2010 ; Vol. 115, No. 19. pp. 3980-3988.
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AB - Activated platelets secrete a negatively charged polymer, polyphosphate (polyP). Here, we explore the interactions of polyP with fibrin(ogen) and its effect on fibrin structure and fibrinolysis. Electrophoretic mobility and binding assays indicate that polyP interacts with fibrinogen and soluble fibrin. Clots formed in the presence of polyP exhibited reduced turbidity and permeability indicative of a tighter fibrin network, but these changes were not related to cross-linking or fibrinopeptide release. Microscopy showed a change in fibrin distribution in clots formed with polyP; with formation of tight aggregates of fibrin fibers interspaced with large pores in contrast to homogenous fiber distribution in control clots. Lysis by tissue plasminogen activator (tPA) and plasminogen or plasmin was delayed in clots formed with polyP and depended on both the activator and polyP concentration. Adding polyP to the clot after fibrin formation or to repolymerizing soluble fibrin did not affect lysis, indicating changes induced by polyP occur at the level of conversion of fibrinogen to fibrin. Surface plasmon resonance showed that the presence of polyP reduced the binding of both plasminogen and tPA to partially lysed fibrin surfaces. These data show that polyP directly influences fibrin architecture and attenuates fibrinolysis through reduced binding of fibrinolytic proteins.

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