Title

Dissecting The Mechanism of Structural Deformation in Transthyretin Amyloidosis

Alternative Title

트렌시스레틴 아밀로이드증에서 나타나는 구조적 변형의 기전 규명

DGIST Authors

Jin-Beom Si ; Jin-Hae Kim ; Jong-Chan Lee

Advisor

김진해

Co-Advisor(s)

Jong-Chan Lee

Issued Date

2026

Awarded Date

2026-08-01

Type

Thesis

Description

Transthyretin, amyloid, aggregation, neurodegenerative diseases

Table Of Contents

I. Introduction 1
1.1 Proteostasis and proteinopathies 1
1.2 Amyloid formation: mechanism, intermediates and pathological consequences 1
1.3 Molecular chaperones and small heat shock protein family 5
II. Structural and Thermodynamic Characterization of Pathogenic Dimeric Transthyretin Variants 7
2.1 Introduction 7
2.1.1 Structural characteristics and functional roles of transthyretin 7
2.1.2 The molecular basis of transthyretin amyloidosis （ATTR） 9
2.2 Materials and methods 12
2.2.1 Recombinant protein production and purification 12
2.2.2 Native-PAGE analysis 13
2.2.3 Analytical SEC and SEC-MALS 13
2.2.4 Circular dichroism （CD） spectroscopy 14
2.2.5 Intrinsic tryptophan fluorescence 14
2.2.6 Thioflavin T （ThT） and turbidity assays 14
2.2.7 [1H-15N] TROSY-HSQC NMR spectroscopy 15
2.2.8 Small-Angle X-ray scattering （SAXS） 15
2.3 Results and discussion 17
2.3.1 Distinct structural characteristics of TTR dimeric species 17
2.3.2 Discrete Biochemical properties of TTR dimers 22
2.3.3 Partially Unfolded or Flexible States of TTR Dimers 25
2.3.4 Conclusion 29
III. HSPB5 Arrests Transthyretin Aggregation by Targeting Early-Stage Misfolded Intermediates 31
3.1 Introduction 31
3.2 Materials and methods 34
3.2.1 Protein expression and purification 34
3.2.2 NMR spectroscopy 34
3.2.3 Turbidity assay and time-of-addition experiments 35
3.2.4 Transmission electron microscopy （TEM） 35
3.2.5 Analytical size-exclusion chromatography and SDS-PAGE 36
3.2.6 Mass photometry 36
3.2.7 Small-Angle X-ray scattering （SAXS） 37
3.3 Results and discussion 38
3.3.1 Conformation selective recognition of destabilized TTR by HSPB5 and its isolated ACD 38
3.3.2 The oligomeric architecture of HSPB5 is required for chaperone activity against TTR aggregation 41
3.3.3 Client engagement drives structural expansion and dynamic oligomeric remodeling of HSPB5 46
3.3.4 HSPB5 selectively targets soluble early-stage aggregation intermediates 50
3.3.5 Conclusions 54
References 58
국문 요약문 65

URI

https://scholar.dgist.ac.kr/handle/20.500.11750/60728
http://dgist.dcollection.net/common/orgView/200001021296

DOI

10.22677/THESIS.200001021296

Degree

Doctor

Department

Department of New Biology

Publisher

DGIST

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