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Identification of novel Fc-interacting proteins involved in the inflammation induced by internalizing anti-DNA antibody
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.author | 김, 민재 | - |
| dc.date.accessioned | 2021-01-06T02:34:50Z | - |
| dc.date.available | 2021-01-06T02:34:50Z | - |
| dc.date.issued | 2020 | - |
| dc.identifier.uri | http://repository.ajou.ac.kr/handle/201003/19243 | - |
| dc.description.abstract | 비공개 | - |
| dc.description.tableofcontents | ABSTRACT i
TABLE OF CONTENTS ii LIST OF FIGURES iii LIST OF TABLES iv I. INTRODUCTION 1 II. MATERIALS AND METHODS 8 A. Cell culture 8 B. Construction of expression vectors and peptide 8 C. Protein preparation using HEK293F cell 9 D. Antibodies 9 E. Lentiviral vector construction 10 F. Establishment of stable cell lines 11 G. Tandem affinity purification (TAP) 11 H. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and in-gel tryptic digestion 12 I. Liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis 13 J. Protein Microarray 13 K. Enzyme-Linked Immunosorbent assay (ELISA) 14 L. Confocal microscopy 14 M. Western blotting 15 N. Immunoprecipitation (IP) 15 O. Quantitative RT-PCR (qRT-PCR) 16 P. In vivo ubiquitination assay 17 Q. Statistical analysis 17 III. RESULTS 18 A. Production of IL-8 and TNF-α induced by cell-internalizable anti-DNA IgG is not mediated by TRIM21-signaling pathway. 18 B. Tandem affinity purification (TAP) is used as a method for the identification of novel Fc-interacting proteins in cell cytosol. 23 C. Extracellular tandem affinity purification (ExTAP) is designed to preserve the structural integrity of IgG-Fc used for identification of novel Fc-receptors in cell cytosol. 28 D. Human IgG Fc-interacting proteins are identified by three independent TAP assays coupled with mass-spectrometry analysis. 33 E. The binding between Fc-interacting proteins and purified IgCw-γκ is confirmed in HEK293T cells. 37 F. The binding between Fc-interacting proteins and internalizing 3D8 IgG is confirmed in HEK293T cells. 41 G. Production of IL-8 and TNF-α response to treatment of 3D8 IgG is reduced by knockdown of Fc-interacting proteins in THP-1 cells. 44 H. Ubiquitination of cell-internalizable 3D8 IgG is not mediated by TRIM21 in THP-1 cells. 48 I. Ubiquitination of 3D8 IgG is increased by TRIM21 knockdown in THP-1 cells. 53 J. β-TrCP2 ubiquitinates 3D8 IgG. 57 IV. DISCUSSION 60 V. CONCLUSION 64 REFERENCES 65 국문요약 72 | - |
| dc.language.iso | en | - |
| dc.title | Identification of novel Fc-interacting proteins involved in the inflammation induced by internalizing anti-DNA antibody | - |
| dc.title.alternative | 세포 유입성 항-DNA 항체 매개의 염증 반응을 조절하는 새로운 Fc 결합 단백질 규명 | - |
| dc.type | Thesis | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000029530 | - |
| dc.subject.keyword | Anti-DNA antibody | - |
| dc.subject.keyword | TRIM21 | - |
| dc.subject.keyword | β-TrCP2 | - |
| dc.subject.keyword | Tandem affinity purification(TAP) | - |
| dc.subject.keyword | 항-DNA 항체 | - |
| dc.subject.keyword | 일렬 친화도 정제 | - |
| dc.description.degree | Doctor | - |
| dc.contributor.department | 대학원 의생명과학과 | - |
| dc.contributor.affiliatedAuthor | 김, 민재 | - |
| dc.date.awarded | 2020 | - |
| dc.type.local | Theses | - |
| dc.citation.date | 2020 | - |
| dc.embargo.liftdate | 9999-12-31 | - |
| dc.embargo.terms | 9999-12-31 | - |
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