Thesis Awital Bar Barroeta
On 24 March 2023 (16:00 hrs) Sanquin researcher Awital Bar Barroeta expects to defend her PhD thesis 'Novel insights into structure and function of Factor XI' at the University of Amsterdam.
Promotores: Prof JCM Meijers PhD and Prof AB Meijer PhD
Venue: Agnietenkapel, Universiteit van Amsterdam and online
Summary
Factor XI (FXI) is a protein in the coagulation cascade. After activation by either factor XIIa (FXIIa) in the intrinsic pathway or by thrombin, activated FXI (FXIa) activates FIX to promote coagulation. FXI provides a supportive role in haemostasis, promoting thrombin generation to ensure fibrin clot integrity and prevent clot degradation. Nevertheless, increased levels of FXI are a risk factor for thrombosis. It has been hypothesised that the prothrombotic role of FXI is associated with its activation by FXIIa, while feedback activation of FXI by thrombin is thought to maintain clot stability in haemostasis. In this thesis we have tried to further explore the structure of FXI and its interactions with proteins to better understand structure-function relationships of FXI and how these relate to FXI function in haemostasis and thrombosis. In this thesis we used hydrogen-deuterium exchange- and crosslinking-mass spectrometry to specify the binding site of HK and FIX on FXI as well as to elucidate conformational changes related to FXI activation. Moreover, the thesis describes the development of a variety of nanobodies against FXI that can be used as tool compounds for the investigation of FXI function. The thesis also reports a time-resolved fluorescence resonance energy transfer (TR-FRET) assay developed to quantify binding between FXI and thrombin-S205A for screening of small molecules with the ability to influence the FXI-thrombin interaction. Together, these findings may aid the development of novel therapeutics regulating FXI function.
Chapters
Chapter 1
General introduction and outline of thesis
Chapter 2
Hydrogen-deuterium exchange mass spectrometry highlights conformational changes induced by factor XI activation and binding of factor IX to factor Xla abstract
Chapter 3
Structures of Factor XI and Prekallikrein bound to domain 6 of High Molecular Weight Kininogen reveals alternate domain 6 conformations and exosites
Chapter 4
Nanobodies against factor XI apple 3 domain inhibit binding of factor IX and reveal a novel binding site for high molecular weight kininogen abstract
Chapter 5
Generation and characterisation of nanobodies targeting FXI
Chapter 6
A FRET-based assay for the quantitation of the thrombin-factor XI interaction abstract
Chapter 7
Thrombin activation of the factor XI dimer is a multi-staged process for each subunit
Chapter 8
General discussion
Download
Download PhD thesis (university repository)
