Suji Kim. (2024). A Study on the Structural & Functional Traits and Effects of Skin-Associated Proteins: Keratin 17 Tail and Erythropoietin. doi: 10.22677/THESIS.200000726587
I. Introduction 1 II. Materials and methods 6 1. In vitro 6 1.1 Plasmids 6 1.2 Protein expression 6 1.3 Inclusion Body (IB) prep 7 1.4 Purification of proteins 7 1.5 Reconstitution of Keratin filament 8 1.6 Assessing keratin filament assemblies and binding 8 1.7 Electron microscope of keratin filaments and measurements of filament width 9 2. In vivo 9 2.1 Cell culture 9 2.2 Sample preparation for skin cell proliferation test 10 2.3 Hematoxylin and eosin (H&E) staining 10 2.4 Western blot 10 2.5 Measurement of the melanin amount 11 3. In slico 12 3.1 Complex structure modeling and analysis 12 4. Statistical analysis 12 III. Results 13 1. The K17 tail domain is essential for filament assembly in the K6/K17 pair 13 1.1 Protein purification for experiment 13 1.2 Sedimentation assay of reconstituted keratin pairs 15 1.3 TEM observations of reconstituted keratin pairs 18 1.4 Self-dimerization of K17T affects keratin filament assembly 19 2. EPO-derived peptides affect skin cell proliferation and melanin synthesis 22 2.1 EPOR is expressed in cells that compose the skin 22 2.2 Designing EPO-derived peptides to reduce side effects and modeling their binding structures to EPOR 23 2.3 Effect of EPO-derived peptides on skin cell proliferation and migration 26 2.4 Effects of EPO-derived peptides on melanin synthesis 31 IV. Discussion 34 1. The K17 tail domain is essential for filament assembly in the K6/K17 pair 34 2. EPO-derived peptides affect skin cell proliferation and melanin synthesis 36 V. Conclusion 37 VI. References 38
