Platelet protein disulfide isomerase promotes glycoprotein ib&-mediated platelet-neutrophil interactions under thromboinflammatory conditions

Jing Li, Kyungho Kim, Si Yeon Jeong, Joyce Chiu, Bei Xiong, Pavel A. Petukhov, Xiangrong Dai, Xiaoyi Li, Robert K. Andrews, Xiaoping Du, Philip J. Hogg, Jaehyung Cho

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Background: Platelet-neutrophil interactions contribute to vascular occlusion and tissue damage in thromboinflammatory disease. Platelet glycoprotein Ib& (GPIb&), a key receptor for the cell-cell interaction, is believed to be constitutively active for ligand binding. Here, we established the role of platelet-derived protein disulfide isomerase (PDI) in reducing the allosteric disulfide bonds in GPIb& and enhancing the ligand-binding activity under thromboinflammatory conditions. Methods: Bioinformatic analysis identified 2 potential allosteric disulfide bonds in GPIb&. Agglutination assays, flow cytometry, surface plasmon resonance analysis, a protein-protein docking model, proximity ligation assays, and mass spectrometry were used to demonstrate a direct interaction between PDI and GPIb& and to determine a role for PDI in regulating GPIb& function and platelet-neutrophil interactions. Also, real-Time microscopy and animal disease models were used to study the pathophysiological role of PDI-GPIb& signaling under thromboinflammatory conditions. Results: Deletion or inhibition of platelet PDI significantly reduced GPIb&-mediated platelet agglutination. Studies using PDI-null platelets and recombinant PDI or Anfibatide, a clinical-stage GPIb& inhibitor, revealed that the oxidoreductase activity of platelet surface-bound PDI was required for the ligand-binding function of GPIb&. PDI directly bound to the extracellular domain of GPIb& on the platelet surface and reduced the Cys4-Cys17 and Cys209-Cys248 disulfide bonds. Real-Time microscopy with platelet-specific PDI conditional knockout and sickle cell disease mice demonstrated that PDI-regulated GPIb& function was essential for platelet-neutrophil interactions and vascular occlusion under thromboinflammatory conditions. Studies using a mouse model of ischemia/reperfusion-induced stroke indicated that PDI-GPIb& signaling played a crucial role in tissue damage. Conclusions: Our results demonstrate that PDI-facilitated cleavage of the allosteric disulfide bonds tightly regulates GPIb& function, promoting platelet-neutrophil interactions, vascular occlusion, and tissue damage under thromboinflammatory conditions.

Original languageEnglish
Pages (from-to)1300-1319
Number of pages20
JournalCirculation
Volume139
Issue number10
DOIs
Publication statusPublished - 5 Mar 2019

Keywords

  • blood platelets
  • glycoprotein Ibalpha
  • inflammation
  • neutrophils
  • protein disulfide isomerase

Cite this

Li, Jing ; Kim, Kyungho ; Jeong, Si Yeon ; Chiu, Joyce ; Xiong, Bei ; Petukhov, Pavel A. ; Dai, Xiangrong ; Li, Xiaoyi ; Andrews, Robert K. ; Du, Xiaoping ; Hogg, Philip J. ; Cho, Jaehyung. / Platelet protein disulfide isomerase promotes glycoprotein ib&-mediated platelet-neutrophil interactions under thromboinflammatory conditions. In: Circulation. 2019 ; Vol. 139, No. 10. pp. 1300-1319.
@article{d9541884199241f8ab178f8792bebe55,
title = "Platelet protein disulfide isomerase promotes glycoprotein ib&-mediated platelet-neutrophil interactions under thromboinflammatory conditions",
abstract = "Background: Platelet-neutrophil interactions contribute to vascular occlusion and tissue damage in thromboinflammatory disease. Platelet glycoprotein Ib& (GPIb&), a key receptor for the cell-cell interaction, is believed to be constitutively active for ligand binding. Here, we established the role of platelet-derived protein disulfide isomerase (PDI) in reducing the allosteric disulfide bonds in GPIb& and enhancing the ligand-binding activity under thromboinflammatory conditions. Methods: Bioinformatic analysis identified 2 potential allosteric disulfide bonds in GPIb&. Agglutination assays, flow cytometry, surface plasmon resonance analysis, a protein-protein docking model, proximity ligation assays, and mass spectrometry were used to demonstrate a direct interaction between PDI and GPIb& and to determine a role for PDI in regulating GPIb& function and platelet-neutrophil interactions. Also, real-Time microscopy and animal disease models were used to study the pathophysiological role of PDI-GPIb& signaling under thromboinflammatory conditions. Results: Deletion or inhibition of platelet PDI significantly reduced GPIb&-mediated platelet agglutination. Studies using PDI-null platelets and recombinant PDI or Anfibatide, a clinical-stage GPIb& inhibitor, revealed that the oxidoreductase activity of platelet surface-bound PDI was required for the ligand-binding function of GPIb&. PDI directly bound to the extracellular domain of GPIb& on the platelet surface and reduced the Cys4-Cys17 and Cys209-Cys248 disulfide bonds. Real-Time microscopy with platelet-specific PDI conditional knockout and sickle cell disease mice demonstrated that PDI-regulated GPIb& function was essential for platelet-neutrophil interactions and vascular occlusion under thromboinflammatory conditions. Studies using a mouse model of ischemia/reperfusion-induced stroke indicated that PDI-GPIb& signaling played a crucial role in tissue damage. Conclusions: Our results demonstrate that PDI-facilitated cleavage of the allosteric disulfide bonds tightly regulates GPIb& function, promoting platelet-neutrophil interactions, vascular occlusion, and tissue damage under thromboinflammatory conditions.",
keywords = "blood platelets, glycoprotein Ibalpha, inflammation, neutrophils, protein disulfide isomerase",
author = "Jing Li and Kyungho Kim and Jeong, {Si Yeon} and Joyce Chiu and Bei Xiong and Petukhov, {Pavel A.} and Xiangrong Dai and Xiaoyi Li and Andrews, {Robert K.} and Xiaoping Du and Hogg, {Philip J.} and Jaehyung Cho",
year = "2019",
month = "3",
day = "5",
doi = "10.1161/CIRCULATIONAHA.118.036323",
language = "English",
volume = "139",
pages = "1300--1319",
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Li, J, Kim, K, Jeong, SY, Chiu, J, Xiong, B, Petukhov, PA, Dai, X, Li, X, Andrews, RK, Du, X, Hogg, PJ & Cho, J 2019, 'Platelet protein disulfide isomerase promotes glycoprotein ib&-mediated platelet-neutrophil interactions under thromboinflammatory conditions' Circulation, vol. 139, no. 10, pp. 1300-1319. https://doi.org/10.1161/CIRCULATIONAHA.118.036323

Platelet protein disulfide isomerase promotes glycoprotein ib&-mediated platelet-neutrophil interactions under thromboinflammatory conditions. / Li, Jing; Kim, Kyungho; Jeong, Si Yeon; Chiu, Joyce; Xiong, Bei; Petukhov, Pavel A.; Dai, Xiangrong; Li, Xiaoyi; Andrews, Robert K.; Du, Xiaoping; Hogg, Philip J.; Cho, Jaehyung.

In: Circulation, Vol. 139, No. 10, 05.03.2019, p. 1300-1319.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Platelet protein disulfide isomerase promotes glycoprotein ib&-mediated platelet-neutrophil interactions under thromboinflammatory conditions

AU - Li, Jing

AU - Kim, Kyungho

AU - Jeong, Si Yeon

AU - Chiu, Joyce

AU - Xiong, Bei

AU - Petukhov, Pavel A.

AU - Dai, Xiangrong

AU - Li, Xiaoyi

AU - Andrews, Robert K.

AU - Du, Xiaoping

AU - Hogg, Philip J.

AU - Cho, Jaehyung

PY - 2019/3/5

Y1 - 2019/3/5

N2 - Background: Platelet-neutrophil interactions contribute to vascular occlusion and tissue damage in thromboinflammatory disease. Platelet glycoprotein Ib& (GPIb&), a key receptor for the cell-cell interaction, is believed to be constitutively active for ligand binding. Here, we established the role of platelet-derived protein disulfide isomerase (PDI) in reducing the allosteric disulfide bonds in GPIb& and enhancing the ligand-binding activity under thromboinflammatory conditions. Methods: Bioinformatic analysis identified 2 potential allosteric disulfide bonds in GPIb&. Agglutination assays, flow cytometry, surface plasmon resonance analysis, a protein-protein docking model, proximity ligation assays, and mass spectrometry were used to demonstrate a direct interaction between PDI and GPIb& and to determine a role for PDI in regulating GPIb& function and platelet-neutrophil interactions. Also, real-Time microscopy and animal disease models were used to study the pathophysiological role of PDI-GPIb& signaling under thromboinflammatory conditions. Results: Deletion or inhibition of platelet PDI significantly reduced GPIb&-mediated platelet agglutination. Studies using PDI-null platelets and recombinant PDI or Anfibatide, a clinical-stage GPIb& inhibitor, revealed that the oxidoreductase activity of platelet surface-bound PDI was required for the ligand-binding function of GPIb&. PDI directly bound to the extracellular domain of GPIb& on the platelet surface and reduced the Cys4-Cys17 and Cys209-Cys248 disulfide bonds. Real-Time microscopy with platelet-specific PDI conditional knockout and sickle cell disease mice demonstrated that PDI-regulated GPIb& function was essential for platelet-neutrophil interactions and vascular occlusion under thromboinflammatory conditions. Studies using a mouse model of ischemia/reperfusion-induced stroke indicated that PDI-GPIb& signaling played a crucial role in tissue damage. Conclusions: Our results demonstrate that PDI-facilitated cleavage of the allosteric disulfide bonds tightly regulates GPIb& function, promoting platelet-neutrophil interactions, vascular occlusion, and tissue damage under thromboinflammatory conditions.

AB - Background: Platelet-neutrophil interactions contribute to vascular occlusion and tissue damage in thromboinflammatory disease. Platelet glycoprotein Ib& (GPIb&), a key receptor for the cell-cell interaction, is believed to be constitutively active for ligand binding. Here, we established the role of platelet-derived protein disulfide isomerase (PDI) in reducing the allosteric disulfide bonds in GPIb& and enhancing the ligand-binding activity under thromboinflammatory conditions. Methods: Bioinformatic analysis identified 2 potential allosteric disulfide bonds in GPIb&. Agglutination assays, flow cytometry, surface plasmon resonance analysis, a protein-protein docking model, proximity ligation assays, and mass spectrometry were used to demonstrate a direct interaction between PDI and GPIb& and to determine a role for PDI in regulating GPIb& function and platelet-neutrophil interactions. Also, real-Time microscopy and animal disease models were used to study the pathophysiological role of PDI-GPIb& signaling under thromboinflammatory conditions. Results: Deletion or inhibition of platelet PDI significantly reduced GPIb&-mediated platelet agglutination. Studies using PDI-null platelets and recombinant PDI or Anfibatide, a clinical-stage GPIb& inhibitor, revealed that the oxidoreductase activity of platelet surface-bound PDI was required for the ligand-binding function of GPIb&. PDI directly bound to the extracellular domain of GPIb& on the platelet surface and reduced the Cys4-Cys17 and Cys209-Cys248 disulfide bonds. Real-Time microscopy with platelet-specific PDI conditional knockout and sickle cell disease mice demonstrated that PDI-regulated GPIb& function was essential for platelet-neutrophil interactions and vascular occlusion under thromboinflammatory conditions. Studies using a mouse model of ischemia/reperfusion-induced stroke indicated that PDI-GPIb& signaling played a crucial role in tissue damage. Conclusions: Our results demonstrate that PDI-facilitated cleavage of the allosteric disulfide bonds tightly regulates GPIb& function, promoting platelet-neutrophil interactions, vascular occlusion, and tissue damage under thromboinflammatory conditions.

KW - blood platelets

KW - glycoprotein Ibalpha

KW - inflammation

KW - neutrophils

KW - protein disulfide isomerase

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U2 - 10.1161/CIRCULATIONAHA.118.036323

DO - 10.1161/CIRCULATIONAHA.118.036323

M3 - Article

VL - 139

SP - 1300

EP - 1319

JO - Circulation

JF - Circulation

SN - 0009-7322

IS - 10

ER -