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Structure-Function Relationship of Keratins in the Context of Epidermolysis Bullosa Simplex and Cancer
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dc.contributor.advisor 이창훈 -
dc.contributor.author Jewoo Kim -
dc.date.accessioned 2025-02-28T21:01:39Z -
dc.date.available 2025-02-28T21:01:39Z -
dc.date.issued 2025 -
dc.identifier.uri http://hdl.handle.net/20.500.11750/58014 -
dc.identifier.uri http://dgist.dcollection.net/common/orgView/200000846149 -
dc.description Keratin 5/14 (K5/K14), Keratin 8/19 (K8/K19), Epidermolysis Bullosa Simplex (EBS), Cancer, GSK3β, 14-3-3, Phosphorylation, Filament assembly, Protein interactions -
dc.description.tableofcontents Ⅰ. Introduction 1
1.1 Keratin5/Keratin14 and Epidermolysis Bullosa Simplex (EBS) 2
1.2 Keratin8/Keratin19 and Cancer 4
II. Methods and Materials 6
2.1 EBS mutant scoring and correlation analysis 6
2.1.1 Homology modeling by MODELLER 6
2.1.2 Aggregation scoring and correlation analysis 8
2.2 Plasmids and Protein expression 8
2.2.1 Mutagenesis and cloning 8
2.2.2 Transformation and protein expression 10
2.3 Protein purification 11
2.3.1 Purification of keratin 11
2.3.2 Purification of GSK3β and GST 12
2.3.3 Purification of 14-3-3 13
2.4 Assembly of Keratin filament 13
2.5 Assessing keratin filament assembly and bundling 14
2.5.1 Sedimentation assay 14
2.5.2 Transmission Electron Microscopy (TEM) 14
2.6 Measurement of material properties of K5/K14 15
2.7 Binding assay of K8/K19 filament with other protein 16
2.7.1 Co-sedimentation assay 16
2.7.2 Nuclear Magnetic Resonance (NMR) 16
III. Results 17
3.1 K5/K14 filaments show altered material properties in EBS mutants 17
3.1.1 Homology modeling 17
3.1.2 Correlation analysis between aggregation score and severity 21
3.1.3 Mutagenesis results 35
3.1.4 Protein purification 36
3.1.5 Sedimentation assay 38
3.1.6 TEM 39
3.1.7 Rheology Analysis 41
3.2 Functional differences between K8/K19 and K8/K19(S35E) 43
3.2.1 Protein purification 43
3.2.2 Sedimentation Assay 47
3.2.3 Comparison of wild-type and mutants (TEM, NMR) 49
3.2.4 Interaction with 14-3-3 (TEM, NMR, co-Sedimentation) 51
3.2.5 Interaction with GSK3β (TEM, NMR, co-Sedimentation) 56
IV. Discussion and Conclusion 61
V. Acknowledgement 65
VI. References 66
Abstract in Korean 70
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dc.format.extent 70 -
dc.language eng -
dc.publisher DGIST -
dc.title Structure-Function Relationship of Keratins in the Context of Epidermolysis Bullosa Simplex and Cancer -
dc.title.alternative 단순 수포성 표피박리증과 암에서 케라틴의 구조-기능 상관관계 연구 -
dc.type Thesis -
dc.identifier.doi 10.22677/THESIS.200000846149 -
dc.description.degree Master -
dc.contributor.department Department of New Biology -
dc.identifier.bibliographicCitation Jewoo Kim. (2025). Structure-Function Relationship of Keratins in the Context of Epidermolysis Bullosa Simplex and Cancer. doi: 10.22677/THESIS.200000846149 -
dc.contributor.coadvisor Jin Hae Kim -
dc.date.awarded 2025-02-01 -
dc.publisher.location Daegu -
dc.description.database dCollection -
dc.citation XT.NM 김74 202502 -
dc.date.accepted 2025-01-20 -
dc.contributor.alternativeDepartment 뉴바이올로지학과 -
dc.subject.keyword Keratin 5/14 (K5/K14), Keratin 8/19 (K8/K19), Epidermolysis Bullosa Simplex (EBS), Cancer, GSK3β, 14-3-3, Phosphorylation, Filament assembly, Protein interactions -
dc.contributor.affiliatedAuthor Jewoo Kim -
dc.contributor.affiliatedAuthor Chang-Hun Lee -
dc.contributor.affiliatedAuthor Jin Hae Kim -
dc.contributor.alternativeName 김제우 -
dc.contributor.alternativeName Chang-Hun Lee -
dc.contributor.alternativeName 김진해 -
dc.rights.embargoReleaseDate 2030-02-28 -
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