Author: Nathaniel Scott Russell
Publisher:
ISBN:
Category :
Languages : en
Pages : 0

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Author: Dara Patrick Dowlatshahi
Publisher:
ISBN:
Category :
Languages : en
Pages :

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The covalent attachment of ubiquitin (Ub), a 76-amino-acid protein, to another protein is a highly occurring and conserved post-translational modification. Ub functions in a plethora of diverse signaling pathways including proteasomal degradation, endocytosis and trafficking of membrane receptors and DNA repair when conjugated to substrate proteins. Ub is unique in that it can also be conjugated to itself to create an assortment of polyubiquitin chains, through either its amino terminus or any of its seven internal lysines, where a large part of its diverse signaling is ascribed to these polyubiquitin chains. Previous work has identified proteins, which specifically recognize these different types of Ub linkages to contain domains or motifs termed Ub binding domains (UBDs). Studies on the various and diverse roles of Ub signaling along with its own diversity of conjugates present on substrates and their recognition by UBDs have uncovered the existence of a Ub syntax or code. The main goal of the work entailed in this thesis was to contribute to decryption of this code by identifying new linkage specific polyubiquitin binding proteins and characterizing their function and mechanism of polyubiquitin recognition. We developed and used a K63 linkage specific polyubiquitin affinity reagent in conjunction with shotgun LC-MS/MS to identify novel polyubiquitin binding proteins. We found that the majority of polyubiquitin dependent identified proteins were previously known Ub binders as well as a few previously unidentified Ub interacting proteins, demonstrating this as a valid approach for the identification of polyubiquitin binding proteins. One such previously unidentified protein we identified was ALIX, a a component of the Endosomal Sorting Complex Required for Transport (ESCRT) machinery known to be involved in Human Immunodeficiency Virus (HIV) budding. We characterized ALIX and found that it binds directly and selectively to K63-linked polyubiquitin via two potential Ub binding sites on a single [alpha]-helical surface within a coiled-coil region, where mutation of these sites impaired retroviral release. Our study demonstrates ALIX as the first example of a K63 chain specific polyubiquitin receptor in the endosomal sorting pathway that supplies evidence for a functional link between K63 polyubiquitin binding and ESCRT function, specifically in retrovirus budding. In summary the findings in our study contribute to the understanding of the role of K63-linked polyubiquitin in the ESCRT pathway. Our work also demonstrates the power of using unbiased affinity capture and tandem mass spectrometry to identify polyubiquitin receptor proteins to further decryption of the Ub code.

Author: Joshua Denis Schnell
Publisher:
ISBN:
Category :
Languages : en
Pages :

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The unicellular eukaryote Saccharomyces cerevisiae can be used as a model system to study ubiquitin-dependent uptake of a signaling receptor. A screen for mutants defective in receptor uptake identified a number of possible candidates genes in this pathway. One of the isolated mutants was characterized further, revealing a novel role for a conserved kinase cascade in regulating endocytosis.

Author: Stephen Joseph Palmieri
Publisher:
ISBN:
Category : Saccharomyces cerevisiae
Languages : en
Pages : 360

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Author:
Publisher:
ISBN:
Category : Dissertations, Academic
Languages : en
Pages : 924

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Author: Thibault Mayor
Publisher: Humana
ISBN: 9781493993710
Category : Science
Languages : en
Pages : 413

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Book Description
“This volume explores numerous techniques used to study the ubiquitin proteasome system. The chapters in this book are organized into five parts and cover topics such as determining the mechanisms of action for E2s, E3s, and DUB enzymes; the latest advances to study the formation of poly-ubiquitin chains as well as their linkage types; the binding partners of proteins in the UPS; methods for structure determination by x-ray crystallography, cryo electron microscopy and SAXS; screening assays to select for degrons or modulators of E3s and DUBs; proteomics approaches in the ubiquitin field and methods to study 26S proteasome function. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Thorough and authoritative, The Ubiquitin Proteasome System: Methods and Protocols is a valuable resource for both experienced and novice scientists who are interested in expanding their knowledge in this field.

Author: Peter Zwickl
Publisher: Springer Science & Business Media
ISBN: 364259414X
Category : Science
Languages : en
Pages : 222

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Book Description
This volume gives an overview of pro tea some-mediated protein degradation and the regulatory role of the ubiquitin system in cellular proteolysis. The first chapter describes the molecular evolution of the proteasome and its associated activators, i. e. , the 20S core, the base and the lid of the 19S cap, and the 11 S regulator. The ensuing chapter gives an overview of the structure and assembly of the 20S proteasome and the regulation of the archaeal proteasome by PAN. The third contribution summarizes our knowledge on the eukaryotic 26S proteasome and its regulation by the 19S regu lator, followed by a chapter devoted to the llS regulator, which elucidates the structural basis for the 11 S-mediated activation of the 20S proteasome. The fifth chapter reviews in detail the role of the proteasome in the immune response. The subsequent chapter of the natural substrates of the gives a comprehensive description proteasome and their recognition by the enzymes of the ubiqui tination machinery. The penultimate chapter rounds up the in formation on intracellular distribution of proteasomes in yeast and mammalian cells, while the last contribution highlights proteasome inhibitors, tools which proved to be very valuable for dissecting the cellular roles of the proteasome and which might turn out to be of pharmacological importance.

Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 2036

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Book Description
Vols. for 1963- include as pt. 2 of the Jan. issue: Medical subject headings.

Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 1320

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Author: J. Robin Harris
Publisher: Springer Nature
ISBN: 3030589714
Category : Science
Languages : en
Pages : 580

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Book Description
This book covers important topics such as the dynamic structure and function of the 26S proteasome, the DNA replication machine: structure and dynamic function and the structural organization and protein–protein interactions in the human adenovirus capsid, to mention but a few. The 18 chapters included here, written by experts in their specific field, are at the forefront of scientific knowledge. The impressive integration of structural data from X-ray crystallography with that from cryo-electron microscopy is apparent throughout the book. In addition, functional aspects are also given a high priority. Chapter 1 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.