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Comprehensive N‐ and O‐glycosylation mapping of human coagulation factor V
Comprehensive N‐ and O‐glycosylation mapping of human coagulation factor V
- Source :
- J Thromb Haemost
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Background/objective Coagulation factor V (FV), a multidomain glycoprotein, is an essential cofactor in the blood clotting cascade. FV deficiency is a rare bleeding disorder that results in poor clotting after an injury or surgery. The only treatment for the disease is infusions of fresh frozen plasma and blood platelets. Glycosylation affects the biological activity, pharmacokinetics, immunogenicity, and in vivo clearance rate of proteins in the plasma. The glycan profile of FV, as well as how it affects the activity, stability, and immunogenicity, remains unknown. Methods In this study, we comprehensively mapped the glycosylation patterns of human plasma-derived FV by combining multienzyme digestion, hydrophilic interaction chromatography enrichment of glycopeptides, and alternated fragmentation mass spectrometry analysis. Results/conclusion A total of 57 unique N-glycopeptides and 51 O-glycopeptides were identified, which were categorized into 40 N-glycan and 17 O-glycan compositions. Such glycosylation details are fundamental for future functional studies and therapeutics development. In addition, the established methodology can be readily applied to analyze glycosylation patterns of proteins with more than 2000 amino acids.
- Subjects :
- chemistry.chemical_classification
Glycosylation
Immunogenicity
Glycopeptides
Factor V
Biological activity
Hematology
030204 cardiovascular system & hematology
Article
Mass Spectrometry
Amino acid
carbohydrates (lipids)
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Biochemistry
chemistry
In vivo
Humans
Platelet
Fresh frozen plasma
Glycoprotein
Glycoproteins
Subjects
Details
- ISSN :
- 15387836
- Volume :
- 18
- Database :
- OpenAIRE
- Journal :
- Journal of Thrombosis and Haemostasis
- Accession number :
- edsair.doi.dedup.....f22294b2b886ad0f39aa4557294735e2
- Full Text :
- https://doi.org/10.1111/jth.14861