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Structure-function of anticoagulant TIX-5, the inhibitor of factor Xa-mediated FV activation. / Maag, Anja; Sharma, Priyanka; Schuijt, Tim J. et al.

In: Journal of thrombosis and haemostasis, Vol. 19, No. 7, 07.2021, p. 1697-1708.

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Maag, Anja ; Sharma, Priyanka ; Schuijt, Tim J. et al. / Structure-function of anticoagulant TIX-5, the inhibitor of factor Xa-mediated FV activation. In: Journal of thrombosis and haemostasis. 2021 ; Vol. 19, No. 7. pp. 1697-1708.

BibTeX

@article{832c380491e24fb490c771b25d073a9f,
title = "Structure-function of anticoagulant TIX-5, the inhibitor of factor Xa-mediated FV activation",
abstract = "Background: The prothrombinase complex consists of factors Xa (FXa) and Va (FVa) on an anionic phospholipid surface and converts prothrombin into thrombin. Both coagulation factors require activation before complex assembly. We recently identified TIX-5, a unique anticoagulant tick protein that specifically inhibits FXa-mediated activation of FV. Because TIX-5 inhibited thrombin generation in blood plasma, it was concluded that FV activation by FXa contributes importantly to coagulation. Objective: We aimed to unravel the structure-function relationships of TIX-5. Method: We used a structure model generated based on homology with the allergen Der F7. Results: Tick inhibitor of factor Xa toward FV was predicted to consist of a single rod formed by several beta sheets wrapped around a central C-terminal alpha helix. By mutagenesis we could show that two hydrophobic loops at one end of the rod mediate the phospholipid binding of TIX-5. On the other end of the rod an FV interaction region was identified on one side, whereas on the other side an EGK sequence was identified that could potentially form a pseudosubstrate of FXa. All three interaction sites were important for the anticoagulant properties of TIX-5 in a tissue factor-initiated thrombin generation assay as well as in the inhibition of FV activation by FXa in a purified system. Conclusion: The structure-function properties of TIX-5 are in perfect agreement with a protein that inhibits the FXa-mediated activation on a phospholipid surface. The present elucidation of the mechanism of action of TIX-5 will aid in deciphering the processes involved in the initiation phase of blood coagulation.",
keywords = "Hemorrhage, anticoagulant, factor V, factor Xa, thrombin",
author = "Anja Maag and Priyanka Sharma and Schuijt, {Tim J.} and Kopatz, {Wil F.} and Dani{\"e}lle Kruijswijk and Marquart, {J. Arnoud} and {van der Poll}, Tom and Hackeng, {Tilman M.} and Nicolaes, {Gerry A. F.} and Meijers, {Joost C. M.} and Bos, {Mettine H. A.} and {van {\textquoteright}t Veer}, Cornelis",
note = "Funding Information: This work was financially supported by the Dutch Thrombosis Foundation (TSN2015‐2) awarded to CvtV and MHAB. The authors gratefully acknowledge Dr. Rodney Camire (Division of Hematology, Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania) for generously providing FV‐B1033 and FV‐B104. Funding Information: This work was financially supported by the Dutch Thrombosis Foundation (TSN2015-2) awarded to CvtV and MHAB. The authors gratefully acknowledge Dr. Rodney Camire (Division of Hematology, Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania) for generously providing FV-B1033 and FV-B104. Publisher Copyright: {\textcopyright} 2021 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals LLC on behalf of International Society on Thrombosis and Haemostasis Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",
year = "2021",
month = jul,
doi = "10.1111/jth.15329",
language = "English",
volume = "19",
pages = "1697--1708",
journal = "Journal of thrombosis and haemostasis",
issn = "1538-7933",
publisher = "Wiley-Blackwell",
number = "7",

}

RIS

TY - JOUR

T1 - Structure-function of anticoagulant TIX-5, the inhibitor of factor Xa-mediated FV activation

AU - Maag, Anja

AU - Sharma, Priyanka

AU - Schuijt, Tim J.

AU - Kopatz, Wil F.

AU - Kruijswijk, Daniëlle

AU - Marquart, J. Arnoud

AU - van der Poll, Tom

AU - Hackeng, Tilman M.

AU - Nicolaes, Gerry A. F.

AU - Meijers, Joost C. M.

AU - Bos, Mettine H. A.

AU - van ’t Veer, Cornelis

N1 - Funding Information: This work was financially supported by the Dutch Thrombosis Foundation (TSN2015‐2) awarded to CvtV and MHAB. The authors gratefully acknowledge Dr. Rodney Camire (Division of Hematology, Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania) for generously providing FV‐B1033 and FV‐B104. Funding Information: This work was financially supported by the Dutch Thrombosis Foundation (TSN2015-2) awarded to CvtV and MHAB. The authors gratefully acknowledge Dr. Rodney Camire (Division of Hematology, Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania) for generously providing FV-B1033 and FV-B104. Publisher Copyright: © 2021 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals LLC on behalf of International Society on Thrombosis and Haemostasis Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/7

Y1 - 2021/7

N2 - Background: The prothrombinase complex consists of factors Xa (FXa) and Va (FVa) on an anionic phospholipid surface and converts prothrombin into thrombin. Both coagulation factors require activation before complex assembly. We recently identified TIX-5, a unique anticoagulant tick protein that specifically inhibits FXa-mediated activation of FV. Because TIX-5 inhibited thrombin generation in blood plasma, it was concluded that FV activation by FXa contributes importantly to coagulation. Objective: We aimed to unravel the structure-function relationships of TIX-5. Method: We used a structure model generated based on homology with the allergen Der F7. Results: Tick inhibitor of factor Xa toward FV was predicted to consist of a single rod formed by several beta sheets wrapped around a central C-terminal alpha helix. By mutagenesis we could show that two hydrophobic loops at one end of the rod mediate the phospholipid binding of TIX-5. On the other end of the rod an FV interaction region was identified on one side, whereas on the other side an EGK sequence was identified that could potentially form a pseudosubstrate of FXa. All three interaction sites were important for the anticoagulant properties of TIX-5 in a tissue factor-initiated thrombin generation assay as well as in the inhibition of FV activation by FXa in a purified system. Conclusion: The structure-function properties of TIX-5 are in perfect agreement with a protein that inhibits the FXa-mediated activation on a phospholipid surface. The present elucidation of the mechanism of action of TIX-5 will aid in deciphering the processes involved in the initiation phase of blood coagulation.

AB - Background: The prothrombinase complex consists of factors Xa (FXa) and Va (FVa) on an anionic phospholipid surface and converts prothrombin into thrombin. Both coagulation factors require activation before complex assembly. We recently identified TIX-5, a unique anticoagulant tick protein that specifically inhibits FXa-mediated activation of FV. Because TIX-5 inhibited thrombin generation in blood plasma, it was concluded that FV activation by FXa contributes importantly to coagulation. Objective: We aimed to unravel the structure-function relationships of TIX-5. Method: We used a structure model generated based on homology with the allergen Der F7. Results: Tick inhibitor of factor Xa toward FV was predicted to consist of a single rod formed by several beta sheets wrapped around a central C-terminal alpha helix. By mutagenesis we could show that two hydrophobic loops at one end of the rod mediate the phospholipid binding of TIX-5. On the other end of the rod an FV interaction region was identified on one side, whereas on the other side an EGK sequence was identified that could potentially form a pseudosubstrate of FXa. All three interaction sites were important for the anticoagulant properties of TIX-5 in a tissue factor-initiated thrombin generation assay as well as in the inhibition of FV activation by FXa in a purified system. Conclusion: The structure-function properties of TIX-5 are in perfect agreement with a protein that inhibits the FXa-mediated activation on a phospholipid surface. The present elucidation of the mechanism of action of TIX-5 will aid in deciphering the processes involved in the initiation phase of blood coagulation.

KW - Hemorrhage

KW - anticoagulant

KW - factor V

KW - factor Xa

KW - thrombin

UR - http://www.scopus.com/inward/record.url?scp=85105242710&partnerID=8YFLogxK

U2 - 10.1111/jth.15329

DO - 10.1111/jth.15329

M3 - Article

C2 - 33829620

VL - 19

SP - 1697

EP - 1708

JO - Journal of thrombosis and haemostasis

JF - Journal of thrombosis and haemostasis

SN - 1538-7933

IS - 7

ER -

ID: 18160722