The objective of this volume is to provide readers with a current view of all aspects of the 'pipeline' that takes protein targets to structures and how these have been optimised. This volume includes chapters describing, in-depth, the individual steps in the Structural Genomics pipeline, as well as less detailed overviews of individual Structural Genomics initiatives. It is the first book of protocols to cover techniques in a new and emerging field.

As research has progressed, the cannabinoid CB 1 and CB 2 receptors have expanded significantly in importance within the neuroscience mainstream. In The Cannabinoid Receptors, leading experts introduce newcomers to the cannabinoid field with chapters covering cannabinoid ligand synthesis and structure activity relationships, the molecular pharmacology of the cannabinoid receptors and the endocannabinoid system, and ultimately, the whole animal pharmacology and therapeutic applications for cannabinoid drugs. Adding to those key topics, the book also examines the current direction of the field with chapters on new putative cannabinoid receptors and challenges for future research. As a part of The Receptors series, this volume highlights its receptor with the most thorough, focused and essential information available.

Comprehensive and cutting-edge, The Cannabinoid Receptors serves as an ideal guidebook to what continues to be a fascinating and vital field.

In recent years remarkable progress has been accomplished with respect to our knowledge about bacterial protein toxins. This refers especially to structural aspects of protein toxins but also holds true for genetics, molecular biology and biochemical mechanisms underlying the action of toxins. This volume covers the very current and exciting aspects of up-to-date bacterial toxicology and comprehensively reviews the most important bacterial protein toxins such as the intracellular acting toxins which exhibit enzyme activity, as well as those toxins that interact with cell plasma membranes by damaging the membranes (pore formation) or stimulating cell receptors (superantigens). This is the most current reference work on these important bacterial protein toxins, which are presented from the point of view of different disciplines such as pharmacology, microbiology, cell biology and protein chemistry.

Quantitative Proteomics by Mass Spectrometry, from the Methods in Molecular Biology series, is a compendium of cutting-edge protocols for quantitative proteomics, and presents the most significant methods used in the field today. The focus on mass spectrometry (MS) is integral, as MS has, and will continue to be, an essential tool in proteomics for studying complex biological systems and human diseases. This volume, written and compiled by leading quantitative proteomic experts, is an indispensable resource in the search for novel biomarkers.
Quantitative Proteomics by Mass Spectrometry presents several innovative MS quantitative procedures, including a variety of methods for introducing isotopic labels and quantifying post-translational modifications. Some of these methods include growing an organism in isotope-enriched media, performing trypsin proteolysis in the presence of 18O-water, reacting protein samples with isotopically labeled reagents, quantifying relative amount of proteins without the use of any isotopic labels. Attention is also given to state-of-the-art techniques for the characterization of the phosphoproteome and tandem MS for detection of inborn errors of metabolism. Specifically, the procedure for determinations of enzymatic activity could be used for large-scale screening of newborns. The protocols in this volume expand both the breadth and depth of readily available methods for quantitative proteomic researchers using MS.

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This book presents broad coverage of the principles and recent developments of sample preparation and fractionation tools in Expression Proteomics in general and two-dimensional electrophoresis (2-DE) in particular. With its unique capacity to resolve thousands of proteins in a single run, 2-DE is still a fundamental research tool for nearly all protein-related scientific projects.

Researchers in structural genomics continue to search for biochemical and cellular functions of proteins as well as the ways in which proteins assemble into functional pathways and networks using either experimental or computational approaches. Based on the experience of leading international experts, Structural Genomics and High Throughput Structural Biology details state-of-the-art analytical and computational methods used to reveal the three-dimensional structure and function of proteins. A historical perspective and a detailed guide to the production of protein material for structural determination, a key step in the process, lay the necessary foundation for discussing the most effective structure determination technologies, such as X-ray crystallography and NMR spectroscopy. Encouraging the study of genes and proteins of unknown structure in order to discover new information about folding, specific structural features, or function, Structural Genomics and High Throughput Structural Biology presents the methods used to interpret the sequences of proteins in a structural context, giving insight into their function. It also explains how to extract information from public data repositories and how to account for variability and accuracy in the quality of this data. The book concludes with a discussion of practical applications of therapeutically driven structural genomics, and presents future directions in the field. Structural Genomics and High Throughput Structural Biology offers a comprehensive guide to the theoretical, technological, and experimental methodologies used to derive structural information from encoded proteins by renowned and world leading scientists in the field.

Volume Two of this two-volume sequence presents a comprehensive overview of protein structure prediction methods and includes protein threading, De novo methods, applications to membrane proteins and protein complexes, structure-based drug design, as well as structure prediction as a systems problem. A series of appendices review the biological and chemical basics related to protein structure, computer science for structural informatics, and prerequisite mathematics and statistics.

Volume One of this two-volume sequence focuses on the basic characterization of known protein structures, and structure prediction from protein sequence information. Eleven chapters survey of the field, covering key topics in modeling, force fields, classification, computational methods, and structure prediction. Each chapter is a self contained review covering definition of the problem and historical perspective; mathematical formulation; computational methods and algorithms; performance results; existing software; strengths, pitfalls, challenges, and future research.

One ofthe major drivers in biological research is the establishment ofstructures and functions of the 50,000 or so proteins in our bodies. Each has a characteristic- dimensional structure, highly "ordered" yet "disordered"! This structure is essential for a protein's function and, significantly, it must be sustained in the competitive and complex environment of the living cell. It is now being recognised that when a cell loses control, proteins can se- assemble into more complex supermolecular structures such as the amyloid fibres and plaques associated with the pathogenesis of prion (CJD) or age-related (Alzheimer's) diseases. This is a pointer to the wider significance of the self-assembling properties of polypeptides. It has been long known that, in silk, polypeptides are assembled into- sheet structures which impart on the material its highly exploitable properties of flexibility combined with high tensile strength. But only now emerging is the recognition that peptides can Self-assemble into a wide variety of non-protein-like structures, including fibrils, fibres, tubules, sheets and monolayers. These are exciting observations and, more so, the potential for materials and medical exploitations is so wide ranging that over 80 scientists from Europe, USA, Japan and Israel. met 1-6 July 1999 in Crete, to discuss the wide-ranging implications of these novel developments. There was a spirit of excitement about the workshop indicative of an important new endeavor. The emerging perception is that of a new class of materials set to become commercially viable early in the 21st century.

Due to the vital biological importance of RNA and proteins functioning together within a cell, a protocol volume describing experimental procedures to study their interactions should find a home in many laboratories. RNA-Protein Interaction Protocols, Second Edition updates, complements, and expands upon the popular first edition by providing a collection of cutting-edge techniques developed or refined in the past few years along with tried-and-true methods. The expert contributors explore the isolation and characterization of RNA-protein complexes, the analysis and measurement of RNA-protein interaction, and related novel techniques and strategies. Written in the highly successful Methods in Molecular Biology (TM) series format, the chapters include brief introductions to the material, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section which highlights tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, RNA-Protein Interaction Protocols, Second Edition is an ideal guide for researchers continuing the study of this all-important biological partnership.

Protein phosphorylation controls many basic cellular processes, such as cell growth, differentiation, migration, metabolism, and cell death, and its study can provide key insights into the signal transduction pathways that are activated in cells in response to different stimuli, such as growth factor stimulation or exposure to toxicants. In Phospho-Proteomics: Methods and Protocols, expert researchers contribute both well-established protocols and some of the newest strategies for the identification and evaluation of protein phosphorylation on Tyr, Ser, and Thr residues, including topics such as 2-dimensional gel electrophoresis and protein phosphorylation, enrichment of phospho-proteins and peptides, quantitative analysis of phosphorylation by labeling and MS analysis, and antibody and kinase arrays. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include brief introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Phospho-Proteomics: Methods and Protocols is an ideal reference for both new and experienced scientists who wish to gain insight into the current developments in the field and to find inspiration to pursue its future.

This book covers elements of both the data-driven comparative modeling approach to structure prediction and also recent attempts to simulate folding using explicit or simplified models. Despite the unsolved mystery of how a protein folds, advances are being made in predicting the interactions of proteins with other molecules. Also rapidly advancing are the methods for solving the inverse folding problem, the problem of finding a sequence to fit a structure.

This book focuses on the various computational methods for prediction, their successes and their limitations, from the perspective of their most well known practitioners.

In the areas of biochemistry and cell biology, characterizations of stability and molecular interactions call for a quantitative approach with a level of precision that matches the fine tuning of these interactions in a living cell. Supporting and up-dating previous Methods in Molecular Biology (TM) volumes, Protein Structure, Stability, and Interactions approaches its subject with a focus on theory and practical applications for both established methods as well as exciting new procedures. The volume presents an overview of many techniques currently used to study protein stability and interactions, including scanning and titration calorimetry, spectroscopic methods, high field NMR, and analytical ultracentrifugation. As a volume of the highly successful Methods in Molecular Biology (TM) series, this work provides the kind of detailed description and implementation advice that is crucial for getting optimal results. Cutting-edge and easy to reference, Protein Structure, Stability, and Interactions is an ideal guide for all scientists interested in biomolecular interactions.

In this book, a select group of researchers has contributed their state-of-the-art methodologies on protein profiling and identification of disease biomarkers in tissues, microdissected cells and body fluids. The book integrates biochemistry, pathology, analytical technology, bioinformatics, and proteome informatics. Experimental approaches are thoroughly detailed and explained through a step-by-step instructional format that ensures successful results.

Intended for Molecular Biologists, Biochemists, and Geneticists interested in manipulating and analyzing the Hedgehog Signaling Pathway, this volume offers various detailed chapters on experimental description, including anecdotal pointers and notes.

This book is specifically about the application of the extremely powerful interaction between the protein avidin or its homologues and the vitamin biotin and some of its homologues. With excellent descriptions of laboratory protocols written by expert researchers, this volume is equally perfect for the student or the professional laboratory scientist.

Over the course of the past decade, there have been remarkable advances in the study of human pathogenic fungi. These developments have taken place throughout a wide range of disciplines, and have come as the result of newly available genome sequences of pathogens such as candida albicans and other model fungi. In Candida Albicans: Methods and Protocols, expert researchers explore these exciting new insights, focusing on the study of medically important fungi and Candida spp in particular. Chapters examine critical aspects of molecular methods, providing information on reporter gene assays, transformation, gene expression in vivo, and methods for large-scale gene disruption. At the same time, the work includes in-depth descriptions of disease models of candidiasis, facts about strain identification, and guidelines on the preparation of samples for proteomic investigations and tandem affinity purification. Composed in the highly successful "Methods in Molecular Biology " series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting edge, Candida Albicans: Methods and Protocols is an invaluable source of methods for investigators in the exhilarating fields of medical and molecular mycology."

Molecular motor proteins produce force for movement in an incredibly wide variety of cellular processes. This volume explores the extreme functional and structural diversity of molecular motors and presents methods relevant to each motor family. In addition, it describes techniques directed at motors that fall outside of the three characterized families: dynamin and F1ATPase.

Post-translational protein modifications by members of the ubiquitin family are widely recognized as important regulatory control systems for a variety of biological pathways. Their influence on eukaryotic cellular metabolism is comparable to that of other modifi- tions such as phosphorylation, acetylation and methylation. The small ubiquitin-related modifier SUMO uses a conjugation and de-conjugation system closely related to that of ubiquitin itself; yet, the functions of the SUMO system are highly diverse and largely independent of the ubiquitin system. SUMO modification controls the activity of tr- scription factors and can influence protein stability, but it also contributes to nucleo-cy- plasmic transport, chromosome segregation and DNA damage repair. As a consequence, the SUMO system pervades virtually all areas of basic molecular and cell biology, and scientists from different backgrounds, including medical researchers, are likely to enco- ter SUMO in the course of their studies. This volume, SUMO Protocols, therefore aims at presenting a collection of methods relevant to SUMO research in order to make these tools available to biochemists, molecular and cell biologists as well as research-oriented clinicians not yet familiar with the system. In contrast to the ubiquitin system, which has been the subject of several reviews and methods collections, no practical compendium entirely devoted to SUMO has been p- lished.

This book surveys the current knowledge concerning the expression and function of stress proteins in different organisms, ranging from prokaryotes to humans. It provides an overview of the diversity and complex evolutionary history of cell stress proteins and describes their function and expression in different eukaryote models. The book will appeal to researchers and scientists in biochemistry, cell biology, microbiology, immunology, and genetics.

This is the first book to examine organelle proteomics in depth.

It begins by introducing the different analytical strategies developed and successfully utilized to study organelle proteomes, and detailing the use of multidimensional liquid chromatography coupled to tandem mass spectrometry for peptide sample analysis. Detailed protocols are provided and a section is devoted to methods enabling a global estimate of the reliability of the protein list assigned to an organelle.

This is a fully up-dated and expanded practical guide to protein structure-function relationships. This important area of research is brought up-to-date by the leading scientists in the field. The compilation of detailed protocols focuses on protein function, proteome research and characterization of pharmaceutical proteins, while following the successful format of the Methods in Molecular Biology series. Comprehensive and cutting edge, the book serves as practical guide for researchers working in the field of protein structure-function relationships and the rapidly growing field of proteomics, as well as scientists in the pharmaceutical industries.

A comprehensive guide to the revolutionary area of systems biology and its application in cell culture engineering, this volume presents an overall picture of the current topics central to structural and functional genomics, proteomics, metabolomics and bioinformatics, including such hot topics as RNAi, metabolic engineering and unfolded protein response. It includes reviews of the cellular response of environmental modulation such as low temperature and osmolarity, critical assessments of the applications of metabolomics and fluxomics approaches, examination of the utility of modulation of key genes and a presentation of a theory of chemical organisation which provides a new view of the system's structure. The clearly written chapters by experts in the field describe methods applicable to investigating the unique facets of cell culture. The book should be of interest to all those working in cell culture development and drug discovery in pharmaceutical and biotechnology companies as well as in academic institutions. It provides an invaluable resource for students and researchers in biotechnology, cell culture, genomics and bioinformatics.

This volume collects new information on the genomics of saprophytic soil Pseudomonas, as well as functions related to genomic islands. It explores life styles in different settings and sheds further insights on the wide metabolic potential of this microbe for the removal of pollutants and production of added-value products. This volume also explores how Pseudomonas responds and reacts to environmental signals, including detection of cell density.

Advanced biotechnologies enable breeders to produce a whole generation of new crops for specialist needs ("designer crops"), including raw materials for the energy, chemical and pharmaceutical industries. This book provides concerns useful to promote an increase of the productivity of crops by using functional genomics (to understand the regulation of plant metabolism at molecular, cellular and whole plants), and the improvement of photosynthetic efficiency (to design new plants with enhanced raw materials percent and recovery). Fundamental thematics have been addressed: metabolic engineering, plant breeding tools, renewable biomass for energy generation, fibres and composites, biopharmaceuticals. The gained know how is relevant to identify bottlenecks in the major production chains and to propose actions for moving these issues forward: in particular to; i) produce new compounds by expressing foreign heterologous genes; ii) modify pathways to influence quality and/or yield of existing indigenous molecules; iii) bioprocess plant or organic waste stream into value-added products. The chapters of this book have been written by experts from all around the world. Consequently, this book is expected to be of great interest to scientists, researchers, farmers, processors and retailers, but also to students, technocrats and planners interested in the progress made with the development of new industrial crops.