Interaction Between the a3 Region of Factor VIII and the TIL'E' Domains of the von Willebrand Factor

Lisbeth Dagil, Kathrin S. Troelsen, Gert Bolt, Lars Thim, Bo Wu, Xin Zhao, Edward G. D. Tuddenham, Thomas E. Nielsen, David Ackland Tanner, Johan H. Faber, Jens Breinholt, Jakob E. Rasmussen*, D. Flemming Hansen

*Corresponding author for this work

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Abstract

The von Willebrand factor (VWF) and coagulation factor VIII (FVIII) are intricately involved in hemostasis. A tight, noncovalent complex between VWF and FVIII prolongs the half-life of FVIII in plasma, and failure to form this complex leads to rapid clearance of FVIII and bleeding diatheses such as hemophilia A and von Willebrand disease (VWD) type 2N. High-resolution insight into the complex between VWF and FVIII has so far been strikingly lacking. This is particularly the case for the flexible a3 region of FVIII, which is imperative for high-affinity binding. Here, a structural and biophysical characterization of the interaction between VWF and FVIII is presented with focus on two of the domains that have been proven pivotal for mediating the interaction, namely the a3 region of FVIII and the TIL'E' domains of VWF. Binding between the FVIII a3 region and VWF TIL'E' was here observed using NMR spectroscopy, where chemical shift changes were localized to two β-sheet regions on the edge of TIL'E' upon FVIII a3 region binding. Isothermal titration calorimetry and NMR spectroscopy were used to characterize the interaction between FVIII and TIL'E' as well as mutants of TIL'E', which further highlights the importance of the β-sheet region of TIL'E' for high-affinity binding. Overall, the results presented provide new insight into the role the FVIII a3 region plays for complex formation between VWF and FVIII and the β-sheet region of TIL'E' is shown to be important for FVIII binding. Thus, the results pave the way for further high-resolution insights into this imperative complex.
Original languageEnglish
JournalBiophysical Journal
Volume117
Issue number3
Pages (from-to)479-489
Number of pages11
ISSN0006-3495
DOIs
Publication statusPublished - 2019

Cite this

Dagil, L., Troelsen, K. S., Bolt, G., Thim, L., Wu, B., Zhao, X., ... Hansen, D. F. (2019). Interaction Between the a3 Region of Factor VIII and the TIL'E' Domains of the von Willebrand Factor. Biophysical Journal, 117(3), 479-489. https://doi.org/10.1016/j.bpj.2019.07.007
Dagil, Lisbeth ; Troelsen, Kathrin S. ; Bolt, Gert ; Thim, Lars ; Wu, Bo ; Zhao, Xin ; Tuddenham, Edward G. D. ; Nielsen, Thomas E. ; Tanner, David Ackland ; Faber, Johan H. ; Breinholt, Jens ; Rasmussen, Jakob E. ; Hansen, D. Flemming. / Interaction Between the a3 Region of Factor VIII and the TIL'E' Domains of the von Willebrand Factor. In: Biophysical Journal. 2019 ; Vol. 117, No. 3. pp. 479-489.
@article{83a76c477b96446b8f6255cdb52053fa,
title = "Interaction Between the a3 Region of Factor VIII and the TIL'E' Domains of the von Willebrand Factor",
abstract = "The von Willebrand factor (VWF) and coagulation factor VIII (FVIII) are intricately involved in hemostasis. A tight, noncovalent complex between VWF and FVIII prolongs the half-life of FVIII in plasma, and failure to form this complex leads to rapid clearance of FVIII and bleeding diatheses such as hemophilia A and von Willebrand disease (VWD) type 2N. High-resolution insight into the complex between VWF and FVIII has so far been strikingly lacking. This is particularly the case for the flexible a3 region of FVIII, which is imperative for high-affinity binding. Here, a structural and biophysical characterization of the interaction between VWF and FVIII is presented with focus on two of the domains that have been proven pivotal for mediating the interaction, namely the a3 region of FVIII and the TIL'E' domains of VWF. Binding between the FVIII a3 region and VWF TIL'E' was here observed using NMR spectroscopy, where chemical shift changes were localized to two β-sheet regions on the edge of TIL'E' upon FVIII a3 region binding. Isothermal titration calorimetry and NMR spectroscopy were used to characterize the interaction between FVIII and TIL'E' as well as mutants of TIL'E', which further highlights the importance of the β-sheet region of TIL'E' for high-affinity binding. Overall, the results presented provide new insight into the role the FVIII a3 region plays for complex formation between VWF and FVIII and the β-sheet region of TIL'E' is shown to be important for FVIII binding. Thus, the results pave the way for further high-resolution insights into this imperative complex.",
author = "Lisbeth Dagil and Troelsen, {Kathrin S.} and Gert Bolt and Lars Thim and Bo Wu and Xin Zhao and Tuddenham, {Edward G. D.} and Nielsen, {Thomas E.} and Tanner, {David Ackland} and Faber, {Johan H.} and Jens Breinholt and Rasmussen, {Jakob E.} and Hansen, {D. Flemming}",
year = "2019",
doi = "10.1016/j.bpj.2019.07.007",
language = "English",
volume = "117",
pages = "479--489",
journal = "Biophysical Journal",
issn = "0006-3495",
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Dagil, L, Troelsen, KS, Bolt, G, Thim, L, Wu, B, Zhao, X, Tuddenham, EGD, Nielsen, TE, Tanner, DA, Faber, JH, Breinholt, J, Rasmussen, JE & Hansen, DF 2019, 'Interaction Between the a3 Region of Factor VIII and the TIL'E' Domains of the von Willebrand Factor', Biophysical Journal, vol. 117, no. 3, pp. 479-489. https://doi.org/10.1016/j.bpj.2019.07.007

Interaction Between the a3 Region of Factor VIII and the TIL'E' Domains of the von Willebrand Factor. / Dagil, Lisbeth; Troelsen, Kathrin S.; Bolt, Gert; Thim, Lars; Wu, Bo; Zhao, Xin; Tuddenham, Edward G. D.; Nielsen, Thomas E.; Tanner, David Ackland; Faber, Johan H.; Breinholt, Jens; Rasmussen, Jakob E.; Hansen, D. Flemming.

In: Biophysical Journal, Vol. 117, No. 3, 2019, p. 479-489.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Interaction Between the a3 Region of Factor VIII and the TIL'E' Domains of the von Willebrand Factor

AU - Dagil, Lisbeth

AU - Troelsen, Kathrin S.

AU - Bolt, Gert

AU - Thim, Lars

AU - Wu, Bo

AU - Zhao, Xin

AU - Tuddenham, Edward G. D.

AU - Nielsen, Thomas E.

AU - Tanner, David Ackland

AU - Faber, Johan H.

AU - Breinholt, Jens

AU - Rasmussen, Jakob E.

AU - Hansen, D. Flemming

PY - 2019

Y1 - 2019

N2 - The von Willebrand factor (VWF) and coagulation factor VIII (FVIII) are intricately involved in hemostasis. A tight, noncovalent complex between VWF and FVIII prolongs the half-life of FVIII in plasma, and failure to form this complex leads to rapid clearance of FVIII and bleeding diatheses such as hemophilia A and von Willebrand disease (VWD) type 2N. High-resolution insight into the complex between VWF and FVIII has so far been strikingly lacking. This is particularly the case for the flexible a3 region of FVIII, which is imperative for high-affinity binding. Here, a structural and biophysical characterization of the interaction between VWF and FVIII is presented with focus on two of the domains that have been proven pivotal for mediating the interaction, namely the a3 region of FVIII and the TIL'E' domains of VWF. Binding between the FVIII a3 region and VWF TIL'E' was here observed using NMR spectroscopy, where chemical shift changes were localized to two β-sheet regions on the edge of TIL'E' upon FVIII a3 region binding. Isothermal titration calorimetry and NMR spectroscopy were used to characterize the interaction between FVIII and TIL'E' as well as mutants of TIL'E', which further highlights the importance of the β-sheet region of TIL'E' for high-affinity binding. Overall, the results presented provide new insight into the role the FVIII a3 region plays for complex formation between VWF and FVIII and the β-sheet region of TIL'E' is shown to be important for FVIII binding. Thus, the results pave the way for further high-resolution insights into this imperative complex.

AB - The von Willebrand factor (VWF) and coagulation factor VIII (FVIII) are intricately involved in hemostasis. A tight, noncovalent complex between VWF and FVIII prolongs the half-life of FVIII in plasma, and failure to form this complex leads to rapid clearance of FVIII and bleeding diatheses such as hemophilia A and von Willebrand disease (VWD) type 2N. High-resolution insight into the complex between VWF and FVIII has so far been strikingly lacking. This is particularly the case for the flexible a3 region of FVIII, which is imperative for high-affinity binding. Here, a structural and biophysical characterization of the interaction between VWF and FVIII is presented with focus on two of the domains that have been proven pivotal for mediating the interaction, namely the a3 region of FVIII and the TIL'E' domains of VWF. Binding between the FVIII a3 region and VWF TIL'E' was here observed using NMR spectroscopy, where chemical shift changes were localized to two β-sheet regions on the edge of TIL'E' upon FVIII a3 region binding. Isothermal titration calorimetry and NMR spectroscopy were used to characterize the interaction between FVIII and TIL'E' as well as mutants of TIL'E', which further highlights the importance of the β-sheet region of TIL'E' for high-affinity binding. Overall, the results presented provide new insight into the role the FVIII a3 region plays for complex formation between VWF and FVIII and the β-sheet region of TIL'E' is shown to be important for FVIII binding. Thus, the results pave the way for further high-resolution insights into this imperative complex.

U2 - 10.1016/j.bpj.2019.07.007

DO - 10.1016/j.bpj.2019.07.007

M3 - Journal article

VL - 117

SP - 479

EP - 489

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 3

ER -