Macromolecular Protein Complexes III: Structure and Function


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.




The Ubiquitin System


Book Description




Plant Photomorphogenesis


Book Description

This book provides detailed protocols for research in plant photomorphogenesis. The collection includes a broad range of topics including assays for shade avoidance responses, assays for light-dependent protein-protein interactions, photobody detection with immunofluorescence and the super-resolution imaging method, protein complex isolation from plants, detection of homodimer and monomer of photoreceptor UVR8 with immunoblotting analysis, assays for seedling greening, procedures for studying skotomorphogenesis, phenotypic study of photomorphogenesis at the seedling stage, expression of Cryptochrome in insect cells, and more. Written for the highly successful Methods in Molecular Biology series, 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. Authoritative and practical, Plant Photomorphogenesis: Methods and Protocols serves as an ideal guide for researchers and students who are new to the field, as well as a stepping stone for experienced researchers to further their skills in this fast-developing field.




Ubiquitin Proteasome System


Book Description

The human ubiquitin proteasome system (UPS) is comprised of nearly 1000 proteins. Although originally identified as a mechanism of protein destruction, the UPS has numerous additional functions and mediates central signaling events in myriad processes involved in both cellular and organismal health and homeostasis. Numerous pathways within the UPS are implicated in disease, ranging from cancer to neurodegenerative diseases such as Parkinson's. The goal of this book is to deliver a collection of synopses of current areas of UPS research that highlights the importance of understanding the biology of the UPS to identify disease-relevant pathways, and the need to elucidate the molecular machinations within the UPS to develop methods for therapeutic modulation of these pathways.




Proteostasis and Disease


Book Description

This book, written by members of the European network PROTEOSTASIS, provides an up-to-date review of the research regarding protein homeostasis in health and disease. With new discoveries contributing to the increasing complexity of this topic, the book offers a detailed overview of the pathways regulating protein homeostasis, including autophagy and the ubiquitin protein family. Following a basic introduction, it explains how defects in protein homeostasis contribute to numerous pathologies, including cancer, neurodegeneration, inflammation and a number of rare diseases. In addition, it discusses, the role of protein homeostasis in cellular development and physiology. Highlighting the latest research in the field of protein homeostasis and its implications for various clinically relevant diseases, the book appeals to researchers and clinicians, while also offering a reference guide for scholars who are new to the field.




The Septin CDCrel-1 [microform] : Protein Associations, Modifications and Effects on Exocytosis


Book Description

SNARE proteins mediate the docking and/or fusion of the vesicle with the plasma membrane. However, it is not clearly understood how this process is regulated. In a search for potential SNARE regulators, we have identified a novel snare interacting protein, the septin CDCrel-1. Septins were first identified as filamentous proteins required for cytokinesis in yeast. However, in mammals little is known about their functions. I show here that cdcrel-1 is predominantly expressed in the brain where it associates with membranes via binding to syntaxin 1A. Wildtype CDCrel-1 transfected into HIT-T15 cells inhibits secretion while mutated forms of CDCrel-1 potentiate secretion, suggesting that cdcrel-1 may be regulating vesicle targeting and/or fusion events. I further map the CDCrel-1 domains important for syntaxin binding and investigate the ability of CDCrel-1 to bind to syntaxin when in various SNARE complexes. CDCrel-1 can bind syntaxin in a SNARE complex, but its binding is occluded by alpha-SNAP. This suggests that CDCrel-1 may act as a novel filamentous element, regulating the delivery and/or fusion of vesicles to the presynaptic membrane through its interaction with syntaxin and the 7S complex. The regulation of filaments may be via post-translational modifications. Indeed we have discovered a novel interaction between SUMO E3 PIAS proteins and CDCrel-1. The conjugation of SUMO to substrates is dependent upon an E1 and E2, whereas specificity is mediated by an E3. Although several SUMO-1 substrates have been characterized, conjugation solely by SUMO-2/3 has not been described. Here I describe the colocalization of CDCrel-1 with SUMO-2 and 3 but not SUMO-1. Transfection of SUMO-2/3 but not SUMO-1 causes a reorganization of CDCrel-1 distribution in CHO cells. Furthermore, CDCrel-1 sequesters the nuclear pool of SUMO-2/3 and of the E2 Ubc9 but not SUMO1 into the cytoplasm. Sumoylation of CDCrel-l is shown in vivo and putative SUMO modification sites on CDCrel-1 are investigated by deletion of lysine residues. These experiments strongly suggest that CDCrel-1 is sumoylated specifically by SUMO-2/3. Sumoylation of CDCrel-1 may therefore play a regulatory role in secretion and septin filament formation. Future work will be aimed at determining the functional significance of SUMO modified CDCrel-1.




Ubiquitin Proteasome System


Book Description

Over the last decade, major advancements in our understanding of the ubiquitin-proteasome system (UPS) have occurred. This book focuses on recent trends in the UPS. The UPS is possibly the most complex of all intracellular pathways as close to 7% of all genes in the human genome make up part of the UPS. This complex system serves as an essential role in intracellular protein degradation, and because of its critical function, improper functioning of the UPS is associated with nearly all know diseases, including cancer, cardiovascular disease, and neurological diseases. The proteolytic component of the UPS is the proteasome, a multicatalytic complex found in the nucleus and cytoplasm. Another form of the proteasome, the immunoproteasome, is less abundant than the constitutive proteasome, but is important in immune response and degradation of oxidized proteins, and recent research suggests that it may be important in longevity. The articles in this book discuss recent findings which indicate that mutations in proteins involved with the UPS are associated with genetic diseases such as familial dilated cardiomyopathy, Nakajo syndrome, and spinal muscular atrophy (X-linked). Some chapters also discuss recent results which suggest that the UPS is heavily regulated by post-translational modifications such as phosphorylation, acetylation, and methylation. The UPS is also heavily regulated by ubiquitination itself. This book contains a research article using PubMed bibliometric data to present current research trends in the UPS. Articles are written so that no one tissue is emphasized to allow readers from any discipline to benefit from this information.




SUMOylation and Ubiquitination


Book Description

Most proteins undergo post-translational modifications altering physical and chemical properties, folding, conformation distribution, stability, activity and function. Ubiquitin and SUMOs are related small proteins that are members of the large ubiquitin superfamily of post-translational modifiers. Written by highly respected leaders in their fields under the expert guidance of the editor, this volume covers the principles of ubiquitination and SUMOylation, presents detailed reviews of current and emerging concepts and highlights new advances in all areas of SUMOylation and ubiquitination. Topics of note include: the ubiquitin superfamily, the ubiquitin toolbox, onco viral exploitation of the SUMO system, small molecule modulators of desumoylation, mass spectrometry, global proteomic profiling of SUMO and ubiquitin, biotin-based approaches, genetic screening, SUMOylation networks in humans, targets for ubiquitin ligases, regulation of p53, protein homeostasis, miRNAs, DNA replication, DNA damage response, telomere biology, intracellular trafficking, regulation of angiogenesis, brain ischemia, autophagy, assembly and activity, antiviral defense, HIV infection, amyloid and amyloid-like proteins, plant immunity. This comprehensive and up-to-date book is the definitive reference volume on all aspects of SUMOylation and ubiquitination and is an essential acquisition for anyone involved in this area of biology.




Plant Meiosis


Book Description

Meiosis is one of the most critical processes in eukaryotes, required for continuation of species and generation of new variation. In plants, meiotic recombination is by far the most important source of genetic variation. In Plant Meiosis: Methods and Protocols, expert researchers in the field detail methods for molecular cytogenetics and chromosome analysis in plants. These state-of -the-art protocols allow studying the organization and behavior of the genetic material in a wide range of both model and crop species. Written in the highly successful Methods in Molecular BiologyTMseries format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Plant Meiosis: Methods and Protocols provides and extensive list of protocols developed and used in a number of laboratories at the cutting edge of meiosis and chromosome research.




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