Study the interaction between plasma Reelin and platelet
| Autor: | Tzu Hsuan Chen, 陳子軒 |
|---|---|
| Rok vydání: | 2012 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 101 The interaction between plasma coagulation proteins and platelet negative charge surface plays an essential role in effective thrombin generation. Reelin is a large plasma protein with positive charge C-terminal tail. Reelin deficient mice revealed decreased thrombin generation. Previous study showed Reelin can interact with platelets through the receptor belonging to the low density lipoprotein receptor gene family, but this interaction can’t be inhibited completely by Reelin well-known receptor antagonist. In this study, we further investigate whether membrane lipids are involved in Reelin-to-platelet interaction. First, solid phase membrane lipid strip screening revealed that Reelin binds to phosphatidylserine (PS), phosphatidic acid (PA), cardiolipin, and sulfatide. On the other hand, we found that 420 kDa full-length Reelin is cleaved into N-terminal 310 kDa and C-terminal 110 kDa fragments by activated coagulation factor X (FXa) in previous study, and we successfully expressed recombinant protein of these fragments. Furthermore, using soluble liposome binding assay revealed that the interaction between Reelin and PS is calcium dependent, and the binding capacity of full-length Reelin and C-terminal 110 kDa fragment is more than N-terminal 310 kDa fragment. Platelet binding assay revealed that full-length Reelin and 310 kDa fragment bind to thrombin-activated PS exposure platelets more than resting platelets. At last, we found that Reelin-to-PS interaction is inhibited by Annexin V both in liposome and platelet binding assay. In this study, we first found that Reelin interacts with PS, and the C-terminal region of Reelin plays an important role on this interaction. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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