Removal of the C-terminal domains of ADAMTS13 by activated coagulation factor XI induces platelet adhesion on endothelial cells under flow conditions

Kathleen S. Garland, Stéphanie E. Reitsma, Toshiaki Shirai, Jevgenia Zilberman-Rudenko, Erik Tucker, David Gailani, Andras Gruber, Owen McCarty, Cristina Puy

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Platelet recruitment to sites of vascular injury is mediated by von Willebrand factor (VWF). The shear-induced unraveling of ultra-large VWF multimers causes the formation of a "stringlike" conformation, which rapidly recruits platelets from the bloodstream. A disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) regulates this process by cleaving VWF to prevent aberrant platelet adhesion; it is unclear whether the activity of ADAMTS13 itself is regulated. The serine proteases α-thrombin and plasmin have been shown to cleave ADAMTS13. Based on sequence homology, we hypothesized that activated coagulation factor XI (FXIa) would likewise cleave ADAMTS13. Our results show that FXIa cleaves ADAMTS13 at the C-terminal domains, generating a truncated ADAMTS13 with a deletion of part of the thrombospondin type-1 domain and the CUB1-2 domains, while α-thrombin cleaves ADAMTS13 near the CUB1-2 domains and plasmin cleaves ADAMTS13 at the metalloprotease domain and at the C-terminal domain. Using a cell surface immunoassay, we observed that FXIa induced the deletion of the CUB1-2 domains from ADAMTS13 on the surface of endothelial cells. Removal of the C-terminal domain of ADAMTS13 by FXIa or α-thrombin caused an increase in ADAMTS13 activity as measured by a fluorogenic substrate (FRETS) and blocked the ability of ADAMTS13 to cleave VWF on the endothelial cell surface, resulting in persistence of VWF strands and causing an increase in platelet adhesion under flow conditions. We have demonstrated a novel mechanism for coagulation proteinases including FXIa in regulating ADAMTS13 activity and function. This may represent an additional hemostatic function by which FXIa promotes local platelet deposition at sites of vessel injury.

Original languageEnglish (US)
Article number232
JournalFrontiers in Medicine
Volume4
Issue numberDEC
DOIs
StatePublished - Jan 1 2017

Fingerprint

Factor XIa
Factor XI
von Willebrand Factor
Blood Platelets
Endothelial Cells
Thrombin
Thrombospondin 1
Fibrinolysin
Metalloproteases
Disintegrins
Vascular System Injuries
Serine Proteases
Hemostatics
Sequence Homology
Fluorescent Dyes
Immunoassay
Peptide Hydrolases
Wounds and Injuries

Keywords

  • A disintegrin and metalloproteinase with a thrombospondin type 1 motif
  • Coagulation
  • Factor XI
  • Member 13
  • Platelets
  • Von Willebrand factor

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Removal of the C-terminal domains of ADAMTS13 by activated coagulation factor XI induces platelet adhesion on endothelial cells under flow conditions. / Garland, Kathleen S.; Reitsma, Stéphanie E.; Shirai, Toshiaki; Zilberman-Rudenko, Jevgenia; Tucker, Erik; Gailani, David; Gruber, Andras; McCarty, Owen; Puy, Cristina.

In: Frontiers in Medicine, Vol. 4, No. DEC, 232, 01.01.2017.

Research output: Contribution to journalArticle

Garland, Kathleen S. ; Reitsma, Stéphanie E. ; Shirai, Toshiaki ; Zilberman-Rudenko, Jevgenia ; Tucker, Erik ; Gailani, David ; Gruber, Andras ; McCarty, Owen ; Puy, Cristina. / Removal of the C-terminal domains of ADAMTS13 by activated coagulation factor XI induces platelet adhesion on endothelial cells under flow conditions. In: Frontiers in Medicine. 2017 ; Vol. 4, No. DEC.
@article{5a3712340a91411b97606eeaf89031b7,
title = "Removal of the C-terminal domains of ADAMTS13 by activated coagulation factor XI induces platelet adhesion on endothelial cells under flow conditions",
abstract = "Platelet recruitment to sites of vascular injury is mediated by von Willebrand factor (VWF). The shear-induced unraveling of ultra-large VWF multimers causes the formation of a {"}stringlike{"} conformation, which rapidly recruits platelets from the bloodstream. A disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) regulates this process by cleaving VWF to prevent aberrant platelet adhesion; it is unclear whether the activity of ADAMTS13 itself is regulated. The serine proteases α-thrombin and plasmin have been shown to cleave ADAMTS13. Based on sequence homology, we hypothesized that activated coagulation factor XI (FXIa) would likewise cleave ADAMTS13. Our results show that FXIa cleaves ADAMTS13 at the C-terminal domains, generating a truncated ADAMTS13 with a deletion of part of the thrombospondin type-1 domain and the CUB1-2 domains, while α-thrombin cleaves ADAMTS13 near the CUB1-2 domains and plasmin cleaves ADAMTS13 at the metalloprotease domain and at the C-terminal domain. Using a cell surface immunoassay, we observed that FXIa induced the deletion of the CUB1-2 domains from ADAMTS13 on the surface of endothelial cells. Removal of the C-terminal domain of ADAMTS13 by FXIa or α-thrombin caused an increase in ADAMTS13 activity as measured by a fluorogenic substrate (FRETS) and blocked the ability of ADAMTS13 to cleave VWF on the endothelial cell surface, resulting in persistence of VWF strands and causing an increase in platelet adhesion under flow conditions. We have demonstrated a novel mechanism for coagulation proteinases including FXIa in regulating ADAMTS13 activity and function. This may represent an additional hemostatic function by which FXIa promotes local platelet deposition at sites of vessel injury.",
keywords = "A disintegrin and metalloproteinase with a thrombospondin type 1 motif, Coagulation, Factor XI, Member 13, Platelets, Von Willebrand factor",
author = "Garland, {Kathleen S.} and Reitsma, {St{\'e}phanie E.} and Toshiaki Shirai and Jevgenia Zilberman-Rudenko and Erik Tucker and David Gailani and Andras Gruber and Owen McCarty and Cristina Puy",
year = "2017",
month = "1",
day = "1",
doi = "10.3389/fmed.2017.00232",
language = "English (US)",
volume = "4",
journal = "Frontiers in Medicine",
issn = "2296-858X",
publisher = "Frontiers Media S. A.",
number = "DEC",

}

TY - JOUR

T1 - Removal of the C-terminal domains of ADAMTS13 by activated coagulation factor XI induces platelet adhesion on endothelial cells under flow conditions

AU - Garland, Kathleen S.

AU - Reitsma, Stéphanie E.

AU - Shirai, Toshiaki

AU - Zilberman-Rudenko, Jevgenia

AU - Tucker, Erik

AU - Gailani, David

AU - Gruber, Andras

AU - McCarty, Owen

AU - Puy, Cristina

PY - 2017/1/1

Y1 - 2017/1/1

N2 - Platelet recruitment to sites of vascular injury is mediated by von Willebrand factor (VWF). The shear-induced unraveling of ultra-large VWF multimers causes the formation of a "stringlike" conformation, which rapidly recruits platelets from the bloodstream. A disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) regulates this process by cleaving VWF to prevent aberrant platelet adhesion; it is unclear whether the activity of ADAMTS13 itself is regulated. The serine proteases α-thrombin and plasmin have been shown to cleave ADAMTS13. Based on sequence homology, we hypothesized that activated coagulation factor XI (FXIa) would likewise cleave ADAMTS13. Our results show that FXIa cleaves ADAMTS13 at the C-terminal domains, generating a truncated ADAMTS13 with a deletion of part of the thrombospondin type-1 domain and the CUB1-2 domains, while α-thrombin cleaves ADAMTS13 near the CUB1-2 domains and plasmin cleaves ADAMTS13 at the metalloprotease domain and at the C-terminal domain. Using a cell surface immunoassay, we observed that FXIa induced the deletion of the CUB1-2 domains from ADAMTS13 on the surface of endothelial cells. Removal of the C-terminal domain of ADAMTS13 by FXIa or α-thrombin caused an increase in ADAMTS13 activity as measured by a fluorogenic substrate (FRETS) and blocked the ability of ADAMTS13 to cleave VWF on the endothelial cell surface, resulting in persistence of VWF strands and causing an increase in platelet adhesion under flow conditions. We have demonstrated a novel mechanism for coagulation proteinases including FXIa in regulating ADAMTS13 activity and function. This may represent an additional hemostatic function by which FXIa promotes local platelet deposition at sites of vessel injury.

AB - Platelet recruitment to sites of vascular injury is mediated by von Willebrand factor (VWF). The shear-induced unraveling of ultra-large VWF multimers causes the formation of a "stringlike" conformation, which rapidly recruits platelets from the bloodstream. A disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) regulates this process by cleaving VWF to prevent aberrant platelet adhesion; it is unclear whether the activity of ADAMTS13 itself is regulated. The serine proteases α-thrombin and plasmin have been shown to cleave ADAMTS13. Based on sequence homology, we hypothesized that activated coagulation factor XI (FXIa) would likewise cleave ADAMTS13. Our results show that FXIa cleaves ADAMTS13 at the C-terminal domains, generating a truncated ADAMTS13 with a deletion of part of the thrombospondin type-1 domain and the CUB1-2 domains, while α-thrombin cleaves ADAMTS13 near the CUB1-2 domains and plasmin cleaves ADAMTS13 at the metalloprotease domain and at the C-terminal domain. Using a cell surface immunoassay, we observed that FXIa induced the deletion of the CUB1-2 domains from ADAMTS13 on the surface of endothelial cells. Removal of the C-terminal domain of ADAMTS13 by FXIa or α-thrombin caused an increase in ADAMTS13 activity as measured by a fluorogenic substrate (FRETS) and blocked the ability of ADAMTS13 to cleave VWF on the endothelial cell surface, resulting in persistence of VWF strands and causing an increase in platelet adhesion under flow conditions. We have demonstrated a novel mechanism for coagulation proteinases including FXIa in regulating ADAMTS13 activity and function. This may represent an additional hemostatic function by which FXIa promotes local platelet deposition at sites of vessel injury.

KW - A disintegrin and metalloproteinase with a thrombospondin type 1 motif

KW - Coagulation

KW - Factor XI

KW - Member 13

KW - Platelets

KW - Von Willebrand factor

UR - http://www.scopus.com/inward/record.url?scp=85062604573&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85062604573&partnerID=8YFLogxK

U2 - 10.3389/fmed.2017.00232

DO - 10.3389/fmed.2017.00232

M3 - Article

AN - SCOPUS:85062604573

VL - 4

JO - Frontiers in Medicine

JF - Frontiers in Medicine

SN - 2296-858X

IS - DEC

M1 - 232

ER -