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El. knyga: Ubiquitin Chains: Degradation and Beyond

  • Formatas: PDF+DRM
  • Serija: Springer Theses
  • Išleidimo metai: 10-Mar-2015
  • Leidėjas: Springer International Publishing AG
  • Kalba: eng
  • ISBN-13: 9783319149653
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Ubiquitin Chains: Degradation and BeyondDidesnis paveikslėlis
  • Formatas: PDF+DRM
  • Serija: Springer Theses
  • Išleidimo metai: 10-Mar-2015
  • Leidėjas: Springer International Publishing AG
  • Kalba: eng
  • ISBN-13: 9783319149653
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This thesis examines the evidence for regulatory ubiquitination by focusing on A20. It provides an insightful and in-depth evaluation of the current literature by critically examining the evidence of K63-linked regulatory ubiquitination in regulating cell-signalling. It is also the first thesis to directly test the role of regulatory ubiquitination in NF-kB signalingin vivo. The case for regulatory ubiquitination has been to a large extent predicated upon the presumed deubiquitinase activity of A20, long considered a key regulator of inflammatory responses as mice lacking A20 die from multi-organ inflammation and cachexia. The theses reports the creation and characterization of a knock-in mouse that expresses a mutated form of A20 which selectively lacks the deubiquitinase activity. The knock-in mice surprisingly display completely normal NF- B activation with no accompanying inflammatory phenotype. Given that the presumed role of A20 as a deubiquitinase has been used to support the importance of regulatory K63-linked ubiquitination in NF-kB signaling, this study will help focus future research efforts into alternative target pathways that do not depend on K63 ubiquitination. In fact, the work suggests that it might be important to revisit the role of K63-linked polyubiquitination in cell-signalling.Ubiquitin Chains: Degradation and Beyond is essential reading for anyone conducting research in cell-signalling and immunology.

Dr. Arnab De received his PhD from the Department of Microbiology & Immunology at Columbia University. During his PhD, he developed transgenic mice to study the mechanism of action of a critical tumor-suppressor called A20. He is also well known for having developed peptide-based prodrugs as therapeutics for diabetes. His work has been reported by the media, and has resulted in multiple patents and publications in peer reviewed journals. He presented his findings at the American Peptide Symposium and was awarded the Young Investigator’s Award. He is the author of the book entitled Application of Peptide-Based Prodrug Chemistry in Drug Development, with a foreword written by Professor Jean Martinez (Former President, European Peptide Society) and published in the series SpringerBriefs in Pharmaceutical Science & Drug Development. His research interests lie at the intersection of chemistry and medicine. Besides biomedical research, he is also generally interested in public health policy and general scientific outreach.
1 Immune Homeostasis: Activation and Down regulation of NF-κB
1(44)
1.1 Introduction to Immune System
1(5)
1.1.1 Innate Immunity
2(1)
1.1.2 Adaptive Immunity
2(4)
1.2 NF-κB: An Overview
6(5)
1.3 NF-κB: The Canonical Pathway and Non-canonical Pathway
11(2)
1.4 A Brief Overview of the IKK Complex
13(2)
1.5 Ubiquitination and NF-κB
15(3)
1.6 Degradative Ubiquitination in Canonical and Non-canonical NF-κB Signaling
18(1)
1.7 Oligomerization in Activation of IKK
18(5)
1.8 Negative Regulation of NF-κB Signaling
23(1)
1.9 Inhibition of NF-κB Activity by Classical IκB Proteins
24(2)
1.10 Inhibition of NF-κB Activity by Deubiquitinases Upstream of IKK
26(5)
1.11 Perspectives on Inhibitors of NF-κB and Broad Aims of this Thesis
31(14)
References
34(11)
2 Regulation of NF-κB Signaling by Regulatory Ubiquitination: Specific Pathways and A20
45(26)
2.1 Regulatory Ubiquitination and NF-κB
45(1)
2.2 IL-1R/TLR Signaling Pathways
46(2)
2.3 TNF Receptor (TNFR) Signaling Pathway
48(5)
2.4 Deubiquitinases and NF-κB
53(1)
2.5 A20 (TNFAIP3 or TNFα-Induced Protein 3 Gene)
54(9)
2.5.1 Structure and Proposed Mechanism of Action
54(2)
2.5.2 Function of A20
56(3)
2.5.3 Regulation of A20 by Ubiquitin-Binding Proteins and Adaptors
59(1)
2.5.4 The Role of A20 in Autoimmune Diseases and Cancer
60(1)
2.5.5 Pathogens Modulating A20 Activity
61(1)
2.5.6 Regulation of A20 Deubiquitinase Activity
62(1)
2.6 Conclusion and Perspectives
63(8)
References
63(8)
3 The Deubiquitinase Activity of A20 Is Dispensable for Its Role in NF-κB Signaling
71(26)
3.1 Introduction
71(2)
3.2 Results
73(16)
3.2.1 Generation of A20-OTU Knock-In Mice
73(2)
3.2.2 Characterization of A20-OTU Knock-In Mice
75(1)
3.2.3 Characterizing the Cells of Immune System of C103A Knock-In Mice in the Steady State
76(2)
3.2.4 Activation of Wild-Type and Mutant BMDM and BMDCs in Response to LPS and TNFα In Vitro
78(7)
3.2.5 Response of A20 OTU Knock-In Mice to LPS Shock
85(2)
3.2.6 Similar Activation of NF-κB in BMDMs from Both Wild-Type and Homozygous Mice
87(2)
3.3 Discussion
89(4)
3.4 Materials and Methods
93(2)
3.4.1 Generation of A20C103A Knock-In Mice
93(1)
3.4.2 Cells
94(1)
3.4.3 Biochemical Experiments
94(1)
3.4.4 LPS-Induced Shock
94(1)
3.4.5 Flow Cytometry, ELISA and qRT--PCR
95(1)
References
95(2)
4 Perspectives on Regulatory Ubiquitination
97(4)
References
99(2)
Index 101
Dr. Arnab De received his PhD from the Department of Microbiology & Immunology at Columbia University. During his PhD, he developed transgenic mice to study the mechanism of action of a critical tumor-suppressor called A20. He is also well known for having developed peptide-based prodrugs as therapeutics for diabetes. His work has been reported by the media, and has resulted in multiple patents and publications in peer reviewed journals. He presented his findings at the American Peptide Symposium and was awarded the Young Investigators Award. He is the author of Application of Peptide-Based Prodrug Chemistry in Drug Development, with a foreword written by Professor Jean Martinez (Former President, European Peptide Society) and published in the series SpringerBriefs in Pharmaceutical Science & Drug Development. His research interests lie at the intersection of chemistry and medicine.  Besides biomedical research, he is also generally interested in public health policy and general scientific outreach.
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