A novel MAL/MyD88-inhibitory decoy peptide attenuates TLR-mediated inflammatory response
- 주제(키워드) Toll-like receptor , MAL/TIRAP , MyD88 , decoy peptide , autoimmune disease
- 발행기관 아주대학교
- 지도교수 최상돈
- 발행년도 2020
- 학위수여년월 2020. 8
- 학위명 석사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000030245
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Stimulated Toll-like receptors (TLRs) trigger the signaling pathway and the first event — the recruitments and interactions of various signaling pathway molecules— occurs to defend our body from the invading pathogens. Among these adaptor proteins, MyD88 adaptor-like (MAL) plays a crucial role in mediating and interaction between the Toll/Interleukin-1 Receptor (TIR) domain-containing signaling molecules such as TLR2/4-TIR and MyD88-TIR. Because of the highly conserved amino acid sequence between the TIR domains, we designed a novel decoy peptide derived from MAL-αC helix that interacts with the TIR domain of MyD88 and TLRs. The designed peptide, MAL/MyD88-inhibitory peptide (MIP), was predicted to interact with MAL, MyD88-TIR, and TLR4-TIR domains. To facilitate cellular uptake, cell-penetrating peptides (CPPs) were linked to the N-terminal of MIP. Only MIP2, linked to penetratin (RQIKIWFQNRRMKWKK), substantially suppressed the NF-κB activation, MyD88- and TRIF-dependent TLRs signaling except TLR3. Through the SPR analysis and immunofluorescence, it is investigated that MIP2 binds to MAL, MyD88-TIR, and TLR4-TIR. Collectively, we report that our decoy peptide, MIP2, has a wide range of TLR inhibitory effects and would be effective in easing autoimmune disease symptoms.
more목차
I. INTRODUCTION 1
II. RESULTS 4
1. Screening of peptide-derived MAL and in silico analyses 4
2. MIP2 hinders the LPS-induced TLR4 downstream transduction and attenuates the oxidative stress. 5
3. MIP2 inhibits TLR signaling in human macrophage cell 7
4. MIP2 binds to TIR domain-containing protein 8
III. DISCUSSION 19
IV. MATERIALS & METHODS 22
1. Cell culture and reagents 22
2. Peptide synthesis 22
3. Cell viability analysis 23
4. ELISA analysis 23
5. SEAP activity analysis 24
6. Western blot analysis 24
7. Confocal microscopy 25
8. NO secretion analysis 26
9. Intracellular NO and ROS analysis 26
10. Surface Plasmon Resonance (SPR) 26
V. REFERENCES 28
