The Anti-Aggregating Activity of HspB1 on Neurodegenerative Disease-Associated Proteins
- 주제(키워드) HspB1 , Naked mole rat , , α-synuclein , Huntingtin , Aβ
- 주제(DDC) 570
- 발행기관 아주대학교 일반대학원
- 지도교수 이창한
- 발행년도 2026
- 학위수여년월 2026. 2
- 학위명 석사
- 학과 및 전공 일반대학원 생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000036079
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The Naked mole rat (NMR) has emerged as a unique model for studying aging-related diseases, including neurodegenerative diseases, due to its exceptionally long lifespan, negligible senescence, and constant mortality rate with age. Among rodents, NMR shows notably elevated expression levels of small heat shock proteins (sHsps), which correlate with lifespan and resistance to age-related pathologies. Neurodegenerative diseases such as Parkinson’s, Alzheimer’s, and Huntington’s are linked to misfolding and aggregation of specific proteins such as α-synuclein (α-Syn), Aβ, and huntingtin (Htt), respectively. We hypothesized that sHsps from NMR possess superior anti-aggregation properties compared to other mammalian homologs. Indeed, the NMR sHsp significantly inhibited the aggregation of a thermolabile model protein and amyloid-forming disease-associated proteins in vitro. Furthermore, it effectively reduced the amyloid-forming protein-induced cytotoxicity in a humanized yeast model, outperforming sHsps from other mammals including mice and human. Structurally, the NMR sHsp contains intrinsically disordered N and C terminal domains flanking a conserved α-crystallin domain (ACD). Its N terminal domain exhibits increased hydrophobicity, which can enhance oligomerization and chaperone activity, contributing to its distinct protective function.
more목차
Chapter 1. The Anti-Aggregating Activity of HspB1 on Neurodegenerative Disease-Associated Proteins 1
1. Introduction 2
2. Methods 5
2.1. Yeast growth spotting assay 5
2.2. Observation of foci-tagged green florescence protein (GFP) 5
2.3. Western blot 6
2.4. His tagged protein purification 6
2.5. Aβ40 purification 7
2.6. α-Syn purification 8
2.7. Htt-46Q purification 8
2.8. Thermal aggregation assay 9
2.9. Thioflavin-T assay 9
2.10. Protein thermal shift assay 10
2.11. Phylogenetic Tree 10
2.12. HspB1 oligomer prediction using Swiss model 11
3. Results 12
3.1. NMR HspB1 is effective in reducing cytotoxicity of 103Q in yeast 12
3.2. NMR HspB1 has better activity than other two HspB1s 13
3.3. NMR HspB1 is more effective in inhibiting aggregates compared to other HspB1s 14
3.4. HspB1s prevent amyloid fibril formation of α-Syn and Aβ40 14
3.5. NMR HspB1 has more alanine sequence in N terminal domain 15
3.6. The N terminal domain swapped variant with NMR HspB1's exhibited better efficiency 16
4. Discussion 18
Chapter 2. The Role of sHsp within Genomic Islands in Bacterial Survival 46
1. Introduction 47
2. Methods 49
2.1. Identification of sHsp homolog using Blast 49
2.2. Phylogenetic tree of sHsp 49
2.3. Gene deletion 49
2.4. Heat resistance spotting assay 50
3. Results 51
3.1. Identification and Comparative Analysis of sHsp Homologs 51
3.2. sHsp20gi2 can substitute for sHsp20gi1 52
3.3. A. baumannii with sHsp has strong heat tolerance 52
4. Discussion 54
References 64
국문요약 77
