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Proteomics Mass Spectrometry Methods: Sample Preparation, Protein Digestion, and Research Protocols shares best practices collected across key laboratories and core facilities, taking the reader through key tactics for executing the most usual mass spectrometry experiments. Sections review research making use of MS proteomics experiments, focus on critical sample preparation, cover mammalian cell lines and samples from clinical tissue and biological fluids, discuss subcellular fractionation, provide methods for protein digestion both for in gel and in solution, and delve into key MS proteomics analysis protocols, including label-free LC-MS, TMT and iTRAQ labelled LC-MS, phosphorylation enrichment, ubiquination enrichment, and more. This book is the perfect lab manual for research teams or for use as a new staff training material. Core facility managers may also find it useful for sharing best practices with their staff and researchers. - Explores the most common questions new researchers have - Guides readers to properly design the workflow for successful integration of mass spectrometry into protein biochemical analyses - Provides examples of sample preparation for a number of different materials, mammalian cells, and others

Principles of Genomics and Proteomics is the perfect reference for graduate students and researchers in these areas to understand its principles and execute precise and reproducible experiments. Following an introductory chapter, the book dives into proper research, including genome mapping. Experiments covered in the book span from Sangers Sequencing, Shotgun sequencing, SAGE analysis, DNA footprinting, Gel retardation, ChIP, and protein resolution methods, including PAGE, 2D gel electrophoresis and isoelectric focusing. Biophysical techniques are also described in detail, including ultraviolet and visible light spectroscopy, fluorescence spectroscopy, NMR and X-ray diffraction. A final proteome analysis is dedicated to functional analysis. Other chapters cover applications of omics technologies broadly. This book is the perfect reference for genetics labs around the world. Graduate students will benefit from the structured and detailed coverage of methods and established researchers will benefit from the book for staff training in research and may find it particularly helpful in enhancing reproducibility of experiments. Provides a comprehensive accounting of the technical aspects of genomic and proteomic analysis and applications in living organisms Includes detailed and standardized coverage of basic research methods in areas covered Presents advanced applications of genomics and proteomics that are highlighted with major significance in medical and agricultural fields

Molecular Biology: Structure and Dynamics of Genomes and Proteomes second edition illustrates the essential principles behind the transmission and expression of genetic information at the level of DNA, RNA, and proteins. Emphasis is on the experimental basis of discovery and the most recent advances in the field while presenting a rigorous, yet still concise, summary of the structural mechanisms of molecular biology. Topics new to this edition include the CRISPR-Cas gene editing system, Coronaviruses – structure, genome, vaccine and drug development, and newly recognized mechanisms for transcription termination. The text is written for advanced undergraduate or graduate-level courses in molecular biology.Key Features Highlights the experimental basis of important discoveries in molecular biology Thoroughly updated with new information on gene editing tools, viruses, and transcription mechanisms, termination and antisense Provides learning objectives for each chapter Includes a list of relevant videos from the Internet about the topics covered in the chapter

Proteomics aims to study all the proteins of human and other living systems, as well as their properties to provide an integrated view of cellular processes. The study of proteomics involves the application of rapidly evolving high-throughput technologies and new platforms that are coming forward regularly, providing versatile novel tools for biomedical and pharmaceutical applications. This book provides a detailed understanding of the basics of proteins and proteomics, gel based-proteomics techniques, basics of mass spectrometry and quantitative proteomics, interactomics: basics and applications, and advancements in proteomics. It also covers basic knowledge about sample preparation, mass spectrometry workflow, different chromatography technologies and quantitative proteomics.The text highlights the application and challenges of various high-throughput integrated proteomics technologies capable of fast and accurate screening of thousands of biomolecules, which are found to be very effective for studying disease pathobiology and identification of next-generation biomarkers and potential drug/vaccine targets; and are therefore considered valuable tools for multidisciplinary research. Features Basics of proteins and proteomics techniques In-depth understanding of mass spectrometry and quantitative proteomics An overview of interactomics and its application for translational research Advancement in the field of proteomics and challenges in clinical applications We hope the knowledge gained from reading this book will intrigue and motivate young minds to explore future opportunities in the constantly evolving field of proteomics.

This book covers different omics aspects related to the extracellular matrix (ECM), namely specific omics resources focused on the extracellular matrix (e.g., databases, repositories and atlases), quantitative proteomics applied to specific extracellular matrices (e.g. basement membranes), biological processes such as ECM degradation (degradomics), cell-matrix interactions (adhesomes), signaling pathways, biomarker discovery and diseases, and interactomics (extracellular matrix interaction networks including not only protein-protein but also protein-glycosaminoglycan interactions). The volume also includes recent advances in glycomics and glycobioinformatics applied to proteoglycans and glycosaminoglycans, which are key biological players. The use of omics data to build dynamic models of ECM-regulated biological pathways is addressed, together with the requirement to standardize omic data, which is a prerequisite for the FAIR (Findability, Accessibility, Interoperability, and Reusability) guiding principles for scientific data management.This book will be of great interest to a broad readership from beginners to advanced researchers, who are interested in extracellular matrix omics and will inspire future research topics.

Methods in Kidney Cell Biology, Volume 153, represents state-of-the-art techniques in renal research that are ideal for veterans, graduate students, postdoctoral fellows, and clinical scientists and principal investigators. Topics in the new release include Single glomerular proteomics – a novel method in translational glomerular cell biology, Measurement of cytosolic and intraciliary calcium in live cells, Differentiation of human kidney organoids from pluripotent stem cells, Quantifying autophagic flux in kidney tissue using structured illumination microscopy, the Generation of primary cells from ADPKD and normal human kidneys, ADPKD cell proliferation and Cl-dependent fluid secretion, In vitro cyst formation of ADPKD cells, and much more. - Written by experts in their field who have perfected the methods they write about - Covers a wide range of topics, from state-of-the-art techniques that may require specialized equipment, to tried-and-true classic methods in their most refined form - Includes cutting-edge, recently developed methods

This new edited volume in the Springer Subcellular Biochemistry Series presents a comprehensive, state-of-the-art overview of the proteomics of peroxisomes derived from mammalian, Drosophila, fungal, and plant origin, and contains contributions from leading experts in the field. The development of sensitive proteomics and mass spectrometry technologies, combined with bioinformatics approaches now allow the identification of low-abundance and transient peroxisomal proteins and permits to identify the complete proteome of peroxisomes, with the consequent increase of our knowledge of the metabolic and regulatory networks of these important cellular organelles. The book lines-up with these developments and is organized in four sections including: (i) mass spectrometry-based organelle proteomics; (ii) prediction of peroxisomal proteomes; (iii) analysis of peroxisome proteome interaction networks; and (iv) peroxisomes in relation to other subcellular compartments. The editor Luis A. del Río is Professor ad honorem of the Spanish National Research Council (CSIC) in the Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Biochemistry and Cell & Molecular Biology of Plants, at the Estación Experimental del Zaidín, Granada, Spain. Del Río's research group focuses on the metabolism of reactive oxygen species (ROS), reactive nitrogen species (RNS) and antioxidants in plant peroxisomes, and the ROS- and RNS-dependent role of peroxisomes in plant cell signalling. The editor Michael Schrader is Professor of Cell Biology & Cytopathology in the Department of Biosciences at the University of Exeter, UK. Using mammalian peroxisomes as model organelles, Prof. Schrader and his team aim to unravel the molecular machinery and signalling pathways that mediate and regulate the formation, dynamics and abundance of these medically relevant cellular compartments.

Sabine Stübler compares different proteasome isoforms and subtypes in terms of their transport and active site-related parameters applying an existing computational model. In a second step, the author extends this model to be able to describe the influence of proteasome inhibitors in in vitro experiments. The computational model, which describes the hydrolysis of short fluorogenic peptides by the 20S proteasome, is calibrated to experimental data from different proteasome isoforms using an approximate Bayesian computation approach. The dynamics of proteasome inhibitors are included into the model in order to demonstrate how to modulate the inhibitor's transport parameters for strong or isoform-specific inhibition.

Molecular Biology: Structure and Dynamics of Genomes and Proteomes illustrates the essential principles behind the transmission and expression of genetic information at the level of DNA, RNA, and proteins. This textbook emphasizes the experimental basis of discovery and the most recent advances in the field in presenting a structural, mechanistic understanding of molecular biology that is rigorous, yet concise. The text is written for a one- or two-term advanced undergraduate/graduate-level course in molecular biology.

This comprehensive volume discusses the protease ADAMTS13, summarizing the current status of basic and clinical research. The nine authoritative chapters begin with a historical perspective followed by exploration of the biochemistry and structure-function relationships of ADAMTS13 as well as its normal function in hemostasis (cleavage of von Willebrand factor). Emerging research themes for ADAMTS13 are covered, including its potential role in angiogenesis and other aspects of cell biology. Additional topics include laboratory assays for ADAMTS13, inherited ADAMTS13 deficiency, and acquired ADAMTS13 deficiency. A chapter on related thrombotic microangiopathic (TMA) disorders examines the differences between TMAs associated with ADAMTS13 deficiency and those not associated with ADAMTS13 deficiency. A final chapter reviews the preliminary information on emerging aspects of ADAMTS13, such as the status of recombinant ADAMTS13 products and their potential utility. Comprehensive in its exploration of the ADAMTS13 protease in disease, ADAMTS13: Biology and Disease is a significant resource for clinical hematologists, transfusion medicine physicians, and researchers interested in hemostasis, vascular biology, biochemistry, and metalloproteases.

Principles of Proteomics, Second Edition, provides a concise and user-friendly introduction to the diverse technologies used for the large-scale analysis of proteins, as well as their applications, and their impact in areas such as drug discovery, agriculture, and the fight against disease. Proteomics is a fast-advancing field in which researchers seek to capture all the proteins in the cell and characterize them in ever more detail. Principles of Proteomics has been fully updated to reflect the most recent developments in the field without losing its focus on the underlying principles. With worked examples, case studies profiling both established and emerging technologies, and further reading lists for each chapter, Principles of Proteomics is an ideal introduction for students, researchers and those working in the industry.

This volume documents this unique family of cell surface proteins. Despite masquerading as intractable and difficult to clone and characterize, ENOX proteins have and continue to offer remarkable opportunities for research, commercial development and outside confirmation of therapeutic, diagnostic and new paradigms to help explain complex biological processes.

In view of rapidly growing research in the deregulation of proteases and their impact in human health and diseases, this book will highlight existing and emerging research in this exciting area. In-depth critical state-of-the-art reviews will be written by established investigators on proteases dysfunctions associated with pathogenesis of different diseases that are known to occur due to deregulation of proteolytic systems. Multidisciplinary approaches demonstrating biochemical and signal transduction mechanisms associated with deregulation of proteases leading to manifestation of the diseases will be discussed. The book highlights the roles of both intracellular and extracellular proteases in health and disease.

The role played by structural proteomics in the first decade of the 21st century is equivalent to that played by the Human Genome Project in the last decade of the 20th century. The development of high-throughput technologies that permit the solution of hundreds of 3D structures of individual proteins, protein-protein complexes and protein-drug complexes, just by one laboratory in a single year, will provide a knowledge base which will change the face of structural biology. This will have an immediate influence on medicinal chemistry and molecular pharmacology, as well as an increasing impact on such disciplines as neurobiology, developmental biology, immunology and molecular medicine.This book presents a state-of-the-art overview of the structural proteomics field, ranging from policy issues related to funding and goals, through the high-throughput procedures for protein production, to the solution of the structures of proteins and higher-order entities, via a multidisciplinary approach involving molecular biology, X-ray crystallography, NMR and electron microscopy, as well as bioinformatics analysis. This is the first book to provide such a comprehensive coverage of a rapidly evolving field.

Still the only concise practical guide to laboratory experiments in proteomics, this new edition now also covers DIGE technology and liquid-chromatography, while the troubleshooting section has been considerably extended. Adopting a practical approach, the authors present the relevant techniques and explain the route to successful experimental design and optimal method selection. They cover such electrophoretic techniques as isoelectric focusing, SDS page, 2-D page, and DIGE, as well as liquid-chromatography techniques, such as ion exchange, affinity chromatography and reversed-phase HPLC. Mass-spectrometric techniques include MALDI, ESI, and FT ICR. Generously illustrated, partly in color, the book also features updates of protocols as well as animations illustrating crucial methodological steps on a companion website.

This volume summarizes the new developments that made subcellular proteomics a rapidly expanding area. It examines the different levels of subcellular organization and their specific methodologies. In addition, the book includes coverage of systems biology that deals with the integration of the data derived from these different levels to produce a synthetic description of the cell as a system.

Rapid growth of the “proteomics” field during the past twelve years has c- tributed to significant advances in science. To date, proteomic technologies have been widely applied to examining various kinds of biological materials. Clinical proteomics is the concept of using proteomic techniques to evaluate the proteomes in clinical samples for biomarker discovery and for better understa- ing of normal physiology and pathogenic mechanisms of human diseases. Tra- lating the proteomic information to clinical practice may lead to the ultimate goals of earlier diagnosis, improved therapeutic outcome and successful prev- tion of diseases. Human body fluids are produced, secreted, and/or excreted from various tissues or organs. Major compositions of body fluids are water, organic s- stances and inorganic compounds. These compositions vary in each body fluid, making one's function different from the others. Analyses of protein com- nents in individual fluids would increase current knowledge on the biology and physiology of various organ systems, and on the pathophysiology of diseases, which cause alterations in protein production, secretion, and/or excretion from the affected tissues or organs into the body fluids. Additionally, human body fluids are the main targets and valuable sources for biomarker discovery. As the high-throughput capability and applicability of proteomics for analyzing proteins in the body fluids have been already proven, these desires are most likely achievable by using proteomic technologies.

Recent developments of experimental techniques in cellular and molecular biology have made it possible to understand the molecular biology of male gametogenesis in greater detail. This book focuses on the description of specialized proteins, which are dominantly and/or specifically expressed in germ cells and localized in spermatozoa. There is an urgent need to classify proteins of spermatogenic cells with a view of their functions, and their applications in the regulation of fertility and in understanding infertility. The understanding of structural properties of male germ cell specific proteins can offer vulnerable points for targeted intervention in testis without generalized effects on stages of spermatogenesis. Besides targeted action in male germ cells, sperm specific proteins and polypeptides may also offer potential application in the development of a contraceptive vaccine.

Since the full functionality of any given protein can only be understood in terms of its interaction with other, often regulatory proteins, this unique reference source covers all relevant protein domains, including SH2, SH3, PDZ, WW, PTB, EH, PH and PX. Its user-oriented concept combines broad coverage with easy retrieval of essential information, and includes a special section on Web-based tools and databases covering protein modules and functional peptide motifs. Essential for the study of protein-protein interactions in vivo or in silico, and a prerequisite for successful functional proteomics studies. With a prologue by Sir Tom Blundell.

This is the very first book to focus on this new approach that will eventually aid in developing new diagnostic markers and therapies for controlling and treating heart disease - the number-one killer in the industrialized world. Divided into two parts, the book describes not only the potentials, but also the limitations of these technologies. The editors, both well known within the scientific community, provide new insights into the biochemical and cellular mechanisms of cardiovascular disease, as well as covering the transition into clinical applications. In so doing, they highlight the various strategies and technical aspects so as to assist the growing number of researchers intending to utilize these approaches. The result is an excellent way of educating and informing graduate students, post-doctoral fellows as well as researchers in academia and industry about the latest developments in this area.

As research on the human, animal, plant and microbial genomes matures towards descriptive fullness, the need for understanding the proteome has clearly emerged as the next major endeavor of life sciences. Proteomics - the quantitative analysis of all proteins working in a cell at a specific time and at specific conditions - provides deep insight into the highly organized network of expression, modification and degradation of proteins. Compiled in this book are reviews and research articles which describe the recent advances and perspectives of this new field of research. The articles are grouped into the following sections: - Sample Preparation and Solubilization - Developments in Electrophoresis - Detection and Quantitation - Mass Spectrometry - Proteome Data Analysis and Management - Prokayotes and Yeast - Biological Fluids - Eukaryotic Cells and Tissue - Oncology - Plants Proteomics is a new key for the functional analysis of living systems and of equal importance for basic as well as application oriented research.