Polyphosphate co-localizes with factor XII on plateletbound fibrin and augments its plasminogen activator activity

Joanne L Mitchell, Ausra S. Lionikiene, Georgi Georgiev, Anja Klemmer, Chelsea Brain, Paul Y Kim, Nicola J. Mutch

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Activated factor XII (FXIIa) has plasminogen activator capacity but its relative contribution to fibrinolysis is considered marginal compared to urokinase and tissue plasminogen activator. Polyphosphate (polyP) is released from activated platelets and mediates FXII activation. Here we investigate the contribution of polyP to the plasminogen activator function of αFXIIa. We show that both polyP70, of the chain length found in platelets (60-100-mer) and platelet-derived polyP, significantly augment the plasminogen activation capacity of αFXIIa. PolyP70 stimulated autoactivation of FXII and subsequent plasminogen activation, indicating that once activated, αFXIIa remains bound to polyP70. Indeed, complex formation between
polyP70 and αFXIIa provides protection against autodegradation. Plasminogen activation by βFXIIa was minimal and was not enhanced by polyP70; highlighting the importance of the anion binding site. PolyP70 did not modulate plasmin activity but stimulated activation of Glu- and Lys- forms of plasminogen by αFXIIa. Accordingly, polyP70 was found to bind to FXII, αFXIIa and plasminogen, but not βFXIIa. Fibrin and polyP70 acted synergistically to enhance αFXIIa-mediated plasminogen activation. The plasminogen activator activity of the αFXIIa-polyP70 complex was modulated by C1 inhibitor and histidine rich glycoprotein, but not plasminogen activator inhibitors (PAI-1 and PAI-2). Platelet polyP and FXII were found to colocalize on the activated platelet membrane in a fibrin-dependent manner and decorated fibrin strands extending from platelet aggregates. We show that in the presence of platelet polyP and the downstream substrate fibrin, αFXIIa is a highly efficient and favorable plasminogen activator. Our data is the first to document aprofibrinolytic function of platelet polyP.
Original languageEnglish
Pages (from-to)2834-2845
Number of pages12
JournalBlood
Volume128
Issue number24
Early online date30 Sep 2016
DOIs
Publication statusPublished - 15 Dec 2016

Fingerprint

Factor XII
Polyphosphates
Plasminogen Activators
Platelets
Fibrin
Plasminogen
Blood Platelets
Chemical activation
Plasminogen Activator Inhibitor 1
Factor XIIa
Plasminogen Activator Inhibitor 2
Plasminogen Inactivators
Fibrinolysin
Urokinase-Type Plasminogen Activator
Fibrinolysis
Tissue Plasminogen Activator
Chain length
Anions
Binding Sites
Membranes

Keywords

  • plasminogen
  • factor XII
  • polyphosphate
  • fibrinolysis
  • fibrin
  • platelets

Cite this

Polyphosphate co-localizes with factor XII on plateletbound fibrin and augments its plasminogen activator activity. / Mitchell, Joanne L; Lionikiene, Ausra S.; Georgiev, Georgi; Klemmer, Anja; Brain, Chelsea; Kim, Paul Y; Mutch, Nicola J.

In: Blood, Vol. 128, No. 24, 15.12.2016, p. 2834-2845.

Research output: Contribution to journalArticle

Mitchell, JL, Lionikiene, AS, Georgiev, G, Klemmer, A, Brain, C, Kim, PY & Mutch, NJ 2016, 'Polyphosphate co-localizes with factor XII on plateletbound fibrin and augments its plasminogen activator activity', Blood, vol. 128, no. 24, pp. 2834-2845. https://doi.org/10.1182/blood-2015-10-673285
Mitchell JL, Lionikiene AS, Georgiev G, Klemmer A, Brain C, Kim PY et al. Polyphosphate co-localizes with factor XII on plateletbound fibrin and augments its plasminogen activator activity. Blood. 2016 Dec 15;128(24):2834-2845. https://doi.org/10.1182/blood-2015-10-673285
Mitchell, Joanne L ; Lionikiene, Ausra S. ; Georgiev, Georgi ; Klemmer, Anja ; Brain, Chelsea ; Kim, Paul Y ; Mutch, Nicola J. / Polyphosphate co-localizes with factor XII on plateletbound fibrin and augments its plasminogen activator activity. In: Blood. 2016 ; Vol. 128, No. 24. pp. 2834-2845.
@article{a44e5a4532954ac68db0f082353ad809,
title = "Polyphosphate co-localizes with factor XII on plateletbound fibrin and augments its plasminogen activator activity",
abstract = "Activated factor XII (FXIIa) has plasminogen activator capacity but its relative contribution to fibrinolysis is considered marginal compared to urokinase and tissue plasminogen activator. Polyphosphate (polyP) is released from activated platelets and mediates FXII activation. Here we investigate the contribution of polyP to the plasminogen activator function of αFXIIa. We show that both polyP70, of the chain length found in platelets (60-100-mer) and platelet-derived polyP, significantly augment the plasminogen activation capacity of αFXIIa. PolyP70 stimulated autoactivation of FXII and subsequent plasminogen activation, indicating that once activated, αFXIIa remains bound to polyP70. Indeed, complex formation betweenpolyP70 and αFXIIa provides protection against autodegradation. Plasminogen activation by βFXIIa was minimal and was not enhanced by polyP70; highlighting the importance of the anion binding site. PolyP70 did not modulate plasmin activity but stimulated activation of Glu- and Lys- forms of plasminogen by αFXIIa. Accordingly, polyP70 was found to bind to FXII, αFXIIa and plasminogen, but not βFXIIa. Fibrin and polyP70 acted synergistically to enhance αFXIIa-mediated plasminogen activation. The plasminogen activator activity of the αFXIIa-polyP70 complex was modulated by C1 inhibitor and histidine rich glycoprotein, but not plasminogen activator inhibitors (PAI-1 and PAI-2). Platelet polyP and FXII were found to colocalize on the activated platelet membrane in a fibrin-dependent manner and decorated fibrin strands extending from platelet aggregates. We show that in the presence of platelet polyP and the downstream substrate fibrin, αFXIIa is a highly efficient and favorable plasminogen activator. Our data is the first to document aprofibrinolytic function of platelet polyP.",
keywords = "plasminogen, factor XII, polyphosphate, fibrinolysis, fibrin, platelets",
author = "Mitchell, {Joanne L} and Lionikiene, {Ausra S.} and Georgi Georgiev and Anja Klemmer and Chelsea Brain and Kim, {Paul Y} and Mutch, {Nicola J.}",
note = "This work was supported by grants FS/11/2/28579 (N.J.M. & A.S.L) from the British Heart Foundation and by the University of Aberdeen Development Trust (J.L.M. & N.J.M.). P.Y.K is supported by an Early Career Award and New Investigator Fund from the Hamilton Health Sciences. We are grateful to the following students for contributions to the project, Natasha Walker & Thomas Nolan. We also thank both the Microscopy and Histology Core Facility and the Iain Fraser Cytometry Centre at the University of Aberdeen for excellent advice and use of the facilities. We also thank Dr Jeffrey Weitz from McMaster University, Canada for the kind gift of HRG.",
year = "2016",
month = "12",
day = "15",
doi = "10.1182/blood-2015-10-673285",
language = "English",
volume = "128",
pages = "2834--2845",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "24",

}

TY - JOUR

T1 - Polyphosphate co-localizes with factor XII on plateletbound fibrin and augments its plasminogen activator activity

AU - Mitchell, Joanne L

AU - Lionikiene, Ausra S.

AU - Georgiev, Georgi

AU - Klemmer, Anja

AU - Brain, Chelsea

AU - Kim, Paul Y

AU - Mutch, Nicola J.

N1 - This work was supported by grants FS/11/2/28579 (N.J.M. & A.S.L) from the British Heart Foundation and by the University of Aberdeen Development Trust (J.L.M. & N.J.M.). P.Y.K is supported by an Early Career Award and New Investigator Fund from the Hamilton Health Sciences. We are grateful to the following students for contributions to the project, Natasha Walker & Thomas Nolan. We also thank both the Microscopy and Histology Core Facility and the Iain Fraser Cytometry Centre at the University of Aberdeen for excellent advice and use of the facilities. We also thank Dr Jeffrey Weitz from McMaster University, Canada for the kind gift of HRG.

PY - 2016/12/15

Y1 - 2016/12/15

N2 - Activated factor XII (FXIIa) has plasminogen activator capacity but its relative contribution to fibrinolysis is considered marginal compared to urokinase and tissue plasminogen activator. Polyphosphate (polyP) is released from activated platelets and mediates FXII activation. Here we investigate the contribution of polyP to the plasminogen activator function of αFXIIa. We show that both polyP70, of the chain length found in platelets (60-100-mer) and platelet-derived polyP, significantly augment the plasminogen activation capacity of αFXIIa. PolyP70 stimulated autoactivation of FXII and subsequent plasminogen activation, indicating that once activated, αFXIIa remains bound to polyP70. Indeed, complex formation betweenpolyP70 and αFXIIa provides protection against autodegradation. Plasminogen activation by βFXIIa was minimal and was not enhanced by polyP70; highlighting the importance of the anion binding site. PolyP70 did not modulate plasmin activity but stimulated activation of Glu- and Lys- forms of plasminogen by αFXIIa. Accordingly, polyP70 was found to bind to FXII, αFXIIa and plasminogen, but not βFXIIa. Fibrin and polyP70 acted synergistically to enhance αFXIIa-mediated plasminogen activation. The plasminogen activator activity of the αFXIIa-polyP70 complex was modulated by C1 inhibitor and histidine rich glycoprotein, but not plasminogen activator inhibitors (PAI-1 and PAI-2). Platelet polyP and FXII were found to colocalize on the activated platelet membrane in a fibrin-dependent manner and decorated fibrin strands extending from platelet aggregates. We show that in the presence of platelet polyP and the downstream substrate fibrin, αFXIIa is a highly efficient and favorable plasminogen activator. Our data is the first to document aprofibrinolytic function of platelet polyP.

AB - Activated factor XII (FXIIa) has plasminogen activator capacity but its relative contribution to fibrinolysis is considered marginal compared to urokinase and tissue plasminogen activator. Polyphosphate (polyP) is released from activated platelets and mediates FXII activation. Here we investigate the contribution of polyP to the plasminogen activator function of αFXIIa. We show that both polyP70, of the chain length found in platelets (60-100-mer) and platelet-derived polyP, significantly augment the plasminogen activation capacity of αFXIIa. PolyP70 stimulated autoactivation of FXII and subsequent plasminogen activation, indicating that once activated, αFXIIa remains bound to polyP70. Indeed, complex formation betweenpolyP70 and αFXIIa provides protection against autodegradation. Plasminogen activation by βFXIIa was minimal and was not enhanced by polyP70; highlighting the importance of the anion binding site. PolyP70 did not modulate plasmin activity but stimulated activation of Glu- and Lys- forms of plasminogen by αFXIIa. Accordingly, polyP70 was found to bind to FXII, αFXIIa and plasminogen, but not βFXIIa. Fibrin and polyP70 acted synergistically to enhance αFXIIa-mediated plasminogen activation. The plasminogen activator activity of the αFXIIa-polyP70 complex was modulated by C1 inhibitor and histidine rich glycoprotein, but not plasminogen activator inhibitors (PAI-1 and PAI-2). Platelet polyP and FXII were found to colocalize on the activated platelet membrane in a fibrin-dependent manner and decorated fibrin strands extending from platelet aggregates. We show that in the presence of platelet polyP and the downstream substrate fibrin, αFXIIa is a highly efficient and favorable plasminogen activator. Our data is the first to document aprofibrinolytic function of platelet polyP.

KW - plasminogen

KW - factor XII

KW - polyphosphate

KW - fibrinolysis

KW - fibrin

KW - platelets

U2 - 10.1182/blood-2015-10-673285

DO - 10.1182/blood-2015-10-673285

M3 - Article

C2 - 27694320

VL - 128

SP - 2834

EP - 2845

JO - Blood

JF - Blood

SN - 0006-4971

IS - 24

ER -