The vast majority of theses in this collection are open access and freely available. There are a small number of theses that have access restricted to the WWU campus. For off-campus access to a thesis labeled "Campus Only Access," please log in here with your WWU universal ID, or talk to your librarian about requesting the restricted thesis through interlibrary loan.
Date of Award
Spring 2024
Document Type
Masters Thesis
Department or Program Affiliation
Chemistry
Degree Name
Master of Science (MS)
Department
Chemistry
First Advisor
Spiegel, P. Clint
Second Advisor
Smirnov, Sergey L.
Third Advisor
Anthony-Cahill, Spencer J.
Abstract
Blood coagulation factor VIII (FVIII) is an essential cofactor in the tenase complex of the blood coagulation cascade for the proteolytic activation of factor X (FX) by activated factor IX (FIXa). FVIII binds to the surface of phosphatidylserine containing membranes via the C1 and C2 domains, increasing the efficiency of FX activation by FIXa 200,000 fold. Structural studies have suggested that FVIII membrane binding is mediated through both hydrophobic and electrostatic interactions between the C domains and lipid surfaces. Many proposed FVIII membrane binding models indicate conflicting binding orientations, highlighting the need for continued structural and biophysical characterization of FVIII lipid binding.This thesis reports thermodynamic, kinetic, and structural data on the C1 and C2 domains binding to lipid nanodiscs (ND). Thermodynamic studies showed that the C1 domain’s binding to lipids is enthalpically driven, whereas the C2 domains binding to lipids is entropically driven. Mutational studies were performed on the C2 domain to understand the membrane binding orientation. Mutants K2183A, D2187A, Q2213A, L2251A, and L2252A had minimal impact on lipid binding, whereas M2199A and F2200A had a significant decrease in lipid binding. Nuclear magnetic resonance (NMR) studies of C2 and ND have highlighted residues interacting with lipids. Molecular dynamics simulations on FVIII and ND have demonstrated novel residues interacting with lipids and support our working model of FVIII membrane binding. This study has provided insight into the ongoing investigation of how FVIII binds to platelet membranes to increase FIXa activity.
Type
Text
Keywords
X-ray Crystallography, NMR, Biochemistry, Molecular Dynamics, Structural Biology, Blood Coagulation Factor VIII (FVIII)
Publisher
Western Washington University
OCLC Number
1435634853
Subject – LCSH
Blood coagulation factor VIII; Blood coagulation factors; Glycoproteins--Structure; Blood proteins--Structure; Blood platelets--Activation; Membrane proteins; Membrane lipids; Nanostructures
Format
application/pdf
Genre/Form
masters theses
Language
English
Rights
Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this document for commercial purposes, or for financial gain, shall not be allowed without the author’s written permission.
Recommended Citation
Avery, Nathan G., "Biophysical and Structural Characterization of Blood Coagulation Factor VIII Lipid Binding with Lipid Nanodiscs" (2024). WWU Graduate School Collection. 1280.
https://cedar.wwu.edu/wwuet/1280