The Role of N-Linked Glycosylation on Human Fibrinogen: A Structure-Function Analysis Utilizing Affinity and High-Performance Liquid Chromatography
Author
Daub, Caroline Elizabeth
Abstract
Fibrinogen (Fgn) is a soluble, 3-chain homodimeric glycoprotein found in blood plasma that is involved in forming blood clots. It is the precursor to fibrin, an insoluble protein that forms the structural scaffold of blood clots. Fibrinogen is converted to fibrin when fibrinopeptides A and B are cleaved off fibrinogen by the enzyme thrombin, exposing polymerization sites termed knob ‘A’ and knob ‘B.’ Blood clots are formed when fibrin molecules laterally aggregate through knob:hole interactions. There are four N-linked glycosylation sites on fibrinogen, which have been suggested to influence the conversion of fibrinogen to fibrin. Furthermore, a relationship is suspected between the pattern of N-linked glycosylation and fibrinopeptide release due to physiological changes during pregnancy. The concentration of fibrinogen in blood plasma rises and the sialic acid content on the N-linked glycans are reduced, which put pregnant women higher potential risk for impaired blood clotting patterns that can lead to stroke or heart attack. However, little is known about the link between glycan structure and the efficiency of thrombin processing the fibrinopeptides. In this preliminary study, a fibrinopeptide release assay (FPRA) using high-performance liquid chromatography (HPLC) was developed to quantify the differences in clotting patterns between native fibrinogen and altered N-linked glycan fibrinogen. Additionally, an affinity column purification system was developed to purify fibrinogen from human blood plasma. In the future, the combination of the affinity column purification system and the FPRA can be used to assess the clotting patterns of pregnant patient fibrinogen samples.
Date
2023-05-05
Citation:
APA:
Daub, Caroline Elizabeth.
(May 2023).
The Role of N-Linked Glycosylation on Human Fibrinogen: A Structure-Function Analysis Utilizing Affinity and High-Performance Liquid Chromatography
(Honors Thesis, East Carolina University). Retrieved from the Scholarship.
(http://hdl.handle.net/10342/13087.)
MLA:
Daub, Caroline Elizabeth.
The Role of N-Linked Glycosylation on Human Fibrinogen: A Structure-Function Analysis Utilizing Affinity and High-Performance Liquid Chromatography.
Honors Thesis. East Carolina University,
May 2023. The Scholarship.
http://hdl.handle.net/10342/13087.
May 20, 2024.
Chicago:
Daub, Caroline Elizabeth,
“The Role of N-Linked Glycosylation on Human Fibrinogen: A Structure-Function Analysis Utilizing Affinity and High-Performance Liquid Chromatography”
(Honors Thesis., East Carolina University,
May 2023).
AMA:
Daub, Caroline Elizabeth.
The Role of N-Linked Glycosylation on Human Fibrinogen: A Structure-Function Analysis Utilizing Affinity and High-Performance Liquid Chromatography
[Honors Thesis]. Greenville, NC: East Carolina University;
May 2023.
Collections
Publisher
East Carolina University