Abstract
Background: Factor (F)XI can be activated by proteases, including thrombin and FXIIa. The interactions of these enzymes with FXI are transient in nature and therefore difficult to study. Objectives: To identify the binding interface between thrombin and FXI and understand the dynamics underlying FXI activation. Methods: Crosslinking mass spectrometry was used to localize the binding interface of thrombin on FXI. Molecular dynamics simulations were applied to investigate conformational changes enabling thrombin-mediated FXI activation after binding. The proposed trajectory of activation was examined with nanobody 1C10, which was previously shown to inhibit thrombin-mediated activation of FXI. Results: We identified a binding interface of thrombin located on the light chain of FXI involving residue Pro520. After this initial interaction, FXI undergoes conformational changes driven by binding of thrombin to the apple 1 domain in a secondary step to allow migration toward the FXI cleavage site. The 1C10 binding site on the apple 1 domain supports this proposed trajectory of thrombin. We validated the results with known mutation sites on FXI. As Pro520 is conserved in prekallikrein (PK), we hypothesized and showed that thrombin can bind PK, even though it cannot activate PK. Conclusion: Our investigations show that the activation of FXI is a multistaged procedure. Thrombin first binds to Pro520 in FXI; thereafter, it migrates toward the activation site by engaging the apple 1 domain. This detailed analysis of the interaction between thrombin and FXI paves a way for future interventions for bleeding or thrombosis.
Original language | English |
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Pages (from-to) | 1336-1346 |
Number of pages | 11 |
Journal | Journal of Thrombosis and Haemostasis |
Volume | 22 |
Issue number | 5 |
Early online date | 17 Jan 2024 |
DOIs | |
Publication status | Published - May 2024 |
Bibliographical note
Publisher Copyright:© 2024 The Author(s)
Funding
J.C.M.M. acknowledges that this research was supported by grant 1702 from the Landsteiner Foundation for Blood Research. R.A.S. acknowledges that this work is part of the research program NWO TA with project number 741.018.201,financed by the Dutch Research Council(NWO). R.A.S. further acknowledges funding through the European Union Horizon 2020 program INFRAIA project Epic-XS (Project 823839). P.A. acknowledges the Dutch National Supercomputer, supported by NWO, for the computational resources (grant agreement EINF-894)
Funders | Funder number |
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Landsteiner Foundation for Blood Research | |
Nederlandse Organisatie voor Wetenschappelijk Onderzoek | 741.018.201 |
Horizon 2020 | EINF-894, 823839 |
Keywords
- factor XI
- mass spectrometry
- prekallikrein
- thrombin
- XL-MS