Galectin-1 and Galectin-3 Constitute Novel-Binding Partners for Factor VIII
| Autor: | Barry J. Byrne, Richard O’Kennedy, Teresa M. Brophy, Jamie M. O’Sullivan, Michelle Lavin, Orla Rawley, P. Vince Jenkins, Alain Chion, Kristina Gegenbauer, Roger J. S. Preston, James S. O’Donnell |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
congenital hereditary and neonatal diseases and abnormalities Glycan Glycosylation Galectin 1 Protein Conformation Galectin 3 Galectins CHO Cells Plasma protein binding 030204 cardiovascular system & hematology Transfection law.invention 03 medical and health sciences chemistry.chemical_compound Cricetulus 0302 clinical medicine Von Willebrand factor law hemic and lymphatic diseases Animals Humans Binding site Blood Coagulation Galectin Binding Sites Factor VIII biology Chemistry Blood Proteins Recombinant Proteins 030104 developmental biology Biochemistry biology.protein Recombinant DNA Partial Thromboplastin Time Cardiology and Cardiovascular Medicine Protein Binding |
| Zdroj: | Arteriosclerosis, Thrombosis, and Vascular Biology. 36:855-863 |
| ISSN: | 1524-4636 1079-5642 |
| DOI: | 10.1161/atvbaha.115.306915 |
| Popis: | Objective— Recent studies have demonstrated that galectin-1 (Gal-1) and galectin-3 (Gal-3) can bind von Willebrand factor and directly modulate von Willebrand factor–dependent early thrombus formation in vivo. Because the glycans expressed on human factor VIII (FVIII) are similar to those of von Willebrand factor, we investigated whether galectins might also bind and modulate the activity of FVIII. Approach and Results— Immunosorbant assays and surface plasmon resonance analysis confirmed that Gal-1 and Gal-3 bound purified FVIII with high affinity. Exoglycosidase removal of FVIII N-linked glycans significantly reduced binding to both Gal-1 and Gal-3. Moreover, combined removal of both the N- and O-glycans of FVIII further attenuated Gal-3 binding. Notably, specific digestion of FVIII high-mannose glycans at N239 and N2118 significantly impaired FVIII affinity for Gal-1. Importantly Gal-1, but not Gal-3, bound to free FVIII in the plasma milieu, and significantly inhibited FVIII functional activity. Interestingly, commercial recombinant FVIII (rFVIII) concentrates are manufactured in different cell lines and differ in their glycosylation profiles. Although the biological mechanism has not been defined, recent studies in previously untreated patients with severe hemophilia A reported significant differences in inhibitor development associated with different rFVIII products. Interestingly, Gal-1 and Gal-3 both displayed enhanced affinity for BHK–rFVIII compared with CHO–rFVIII. Furthermore, binding of Gal-1 and Gal-3 to BDD–FVIII was markedly reduced compared with full-length rFVIII. Conclusions— We have identified Gal-1 and Gal-3 as novel-binding partners for human FVIII and demonstrated that Gal-1 binding can influence the procoagulant activity of FVIII. |
| Databáze: | OpenAIRE |
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