Title Page
Contents
ABSTRACT 8
Chapter 1. Introduction 9
Chapter 2. Materials and methods 17
2.1. Gene cloning 17
2.2. Overexpression and purification of human FGF10 19
2.3. Crystallization of human FGF10 21
2.4. Structure determination and refinements 26
2.5. Cellular functional assay of FGF10 and truncated FGF10 26
Chapter 3. Results 27
3.1. Crystal structure of FGF10 alone 27
3.2. Structure comparison between FGF10 alone and in complex with FGFR2b 30
3.3. Receptor-binding induced conformation change of FGF10 30
3.4. Binding interactions between FGF10 α1 and FGFR2b 33
3.5. Specificity between FGF10 and FGFR2b 33
3.6. Conformation-changing α1 part contributes to the cell proliferation activity of FGF10 37
Chapter 4. Conclusion 44
References 48
Abstract (in korean) 52
TABLE 1. Data collections and refinement statistics 28
TABLE 2. Interactions between FGF10 and FGFR2b in complex structure 32
TABLE 3. Solvent accessible area of FGF10 with α1 and without α1 related to the FGFR2b binding 34
TABLE 4. Determined crystal structures of FGFs alone and in complex with FGFRs 35
Figure 1. The Diverse Functions of FGF/FGFR Signaling. 10
Figure 2. Sequence Alignment of Receptor Types B and C. 11
Figure 3. Decoding the Pathways: Unraveling the Intricacies of Classical FGF/FGFR Signaling. 13
Figure 4. Decoding Heparan Sulfate Proteoglycans (HSPGs) in Angiogenesis. 15
Figure 5. The human fibroblast growth factor (FGF) gene family. 16
Figure 6. FGF10 and pET24a Expression Vector Digestion. 18
Figure 7. Two-Step Method for Purification of FGF10. 20
Figure 8. Expression test of FGF10 protein. 22
Figure 9. Purification process of the FGF10 protein. 23
Figure 10. Process of crystallization of FGF10 protein. 24
Figure 11. FGF10 Protein as Determined by Gel Filtration. 25
Figure 12. Superimpose structures of FGF10. 29
Figure 13. Summarizes the topology diagrams of the FGF10 structure. 31
Figure 14. Structure alignment of paracrine FGFs. 36
Figure 15. Cell proliferation assay of FGF10. 38
Figure 16. SDS-PAGE analysis of native FGF10 truncated FGF10. 39
Figure 17. Conformational Changes of FGF1 and FGF9 N-Terminus. 40
Figure 18. FGFs Dimer Conformation. 41
Figure 19. The structural variations in FGF dimers. 42
Figure 20. Insights into Loop Dynamics and N-Terminal Interaction. 43