Senior Project Advisor

Clint Spiegel

Document Type

Project

Publication Date

Spring 2022

Keywords

Blood Coagulation, Factor IX, Refolding Experiment

Abstract

This paper attempts to provide an optimized strategy for the purification, activation, and isolation of blood coagulation Factor IX mutants. The goal of this work is to enable future biochemical and structural studies of Factor IX to a gain a better understanding of the structural-functional role this protein plays in the blood coagulation cascade. The orchestration and amplification of the blood coagulation cascade requires the binding of Factor VIII (FVIII) to an activated platelet surface, where it serves as a cofactor to a serine protease, Factor IX (FIX). Factor IX circulates the bloodstream as a catalytically silent multidomain protein1. Like many other coagulation factors, FIX requires cofactor-triggered and substrate-assisted modulations to become active2. Following activation of FIXa, formation of the FIXa/FVIIIa ‘Xase’ complex is enabled by structural rearrangements in the 99-loop and 60-loop of FIXa that allow for the proteolytic cleavage or turnover of FX to FXa2,3. The 200,000 fold activity enhancement that culminates from Xase formation is responsible for thrombin generation and blood clot formation4. A structural understanding of FIXa/FVIIIa activity and assembly in the Xase complex remains largely unknown and represents a fundamental challenge in the physiological understanding of blood coagulation disorders.

Department

Chemistry

Type

Text

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.

Language

English

Format

application/pdf

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