This book presents pioneering findings on the characterization of cellular regulation and function for three recently identified protein posttranslational modifications (PTMs): lysine malonylation (Kmal), glutarylation (Kglu) and crotonylation (Kcr). It addresses three main topics: (i) Detecting Kmal substrates using a chemical reporter, which provides important information regarding the complex cellular networks modulated by Kmal; (ii) Identifying Kglu as a new histone PTM and assessing the direct impact of histone Kglu on chromatin structure and dynamics; and (iii) Revealing Sirt3's value as a regulating enzyme for histone Kcr dynamics and gene transcription, which opens new avenues for examining the physiological significance of histone Kcr. Taken together, these studies provide information critical to understanding how these protein PTMs are associated with various human diseases, and to identifying therapeutic targets for the dysregulation of these novel protein markers in various human diseases.

The second volume continues to fill the gap in protein review and protocol literature. It does this while summarizing recent achievements in the understanding of the relationships between protein misfoldings, aggregation, and development of protein deposition disorders. The focus of Part B is the molecular basis of differential disorders.

This volume presents established bioinformatics tools and databases for function prediction of proteins. Reflecting the diversity of this active field in bioinformatics, the chapters in this book discuss a variety of tools and resources such as sequence-, structure-, systems-, and interaction-based function prediction methods, tools for functional analysis of metagenomics data, detecting moonlighting-proteins, sub-cellular localization prediction, and pathway and comparative genomics databases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, step-by-step instructions of how to use software and web resources, use cases, and tips on troubleshooting and avoiding known pitfalls. Thorough and cutting-edge, Protein Function Prediction: Methods and Protocols is a valuable and practical guide for using bioinformatics tools for investigating protein function

This book covers the latest findings of a wide variety of viral, prokaryotic and eukaryotic macromolecular protein complexes and builds upon the solid macromolecular foundations established by previous volumes of the Subcellular Biochemistry series. Thus, an almost encyclopaedic coverage of the broad field of protein complex structure and function has been established. The 17 interesting chapters included in this book have been organised into four sections: Soluble Protein Complexes, Membrane Protein Complexes, Fibrous Protein Complexes and Viral Protein Complexes. Significant topics present here are: Fatty Acid Synthase, the Fork Protection Complex, Ribonucleotide Reductase, the Kinetochore, G proteins, the FtsEX Complex, the Kainate Receptor, the Photosystem I-antenna, the Mycobacterial Arabinofuranosyltransferases, the the Bacterial Flagellum, the Actomyosin Complex, Motile Cilia, SLS Collagen Polymorphic Structures, and the Reovirus Capsid and Polymerase. Up-dates/expansion of chapter topics present in earlier volumes are now included in chapters here, e.g., those on Ferritin-like proteins and the Multi-tRNA Synthetase. The book is richly illustrated throughout, the result of an impressive integration of structural data from X-ray crystallography and cryo-electron microscopy. The functional aspects of protein-protein interactions are also given a high priority.

The main purpose of this volume is to provide a focused analysis of the function of the G protein-coupled signaling pathways that operate in the interconnected network of retinal neurons as they detect and encode the information carried by light. The organization of this volume will generally follow the path of signal flow in the retina. First we will describe recent advances in understanding the phototransduction cascade of rod and cone photoreceptors, which use signaling cascade based on the GPCR rhodopsin to transduce incident light into neural activity. Chapters will be devoted to unique specializations of the two major types of photosensitive cells that comprise the predominant input for our spatial and color vision. Subsequently, the mechanisms of synaptic information encoding by retinal ON bipolar cells will be described, where the GPCR mGluR6 plays a fundamental role. Chapters in this section will examine macromolecular organization of the mGluR6 signaling pathway as well as current understanding of its function. The functional characteristics of this signaling mechanism will be explored in detail. Additionally, this section will cover the role of dopamine receptors in modulating signal transmission between photoreceptors and ON-bipolar cells. Finally, chapters will be focused on the output neurons of the inner retina, ganglion cells, where the components of the emerging GPCR melanopsin cascade in intrinsically photosensitive ganglion cells will be detailed. Collectively these mechanisms allow the retina to represent visual space over a wide range of light intensities.

This book explores the broad and diverse biological and physiological impacts of established and newly discovered cyclic di-nucleotide second messenger signaling systems, while also providing descriptions of the intriguing biochemical characteristics of multiple turnover enzymes and receptors. The respective chapters discuss the commonalities and diversity of cyclic di-GMP, cyclic di-AMP and recently discovered cyclic GMP-AMP signaling systems in manifold Gram-negative and Gram-positive bacteria. The global human pathogens Mycobacterium tuberculosis, Vibrio cholerae, Salmonella typhimurium, Escherichia coli and Streptococcus pneumoniae, the facultative human pathogen Pseudomonas aeruginosa, global plant pathogens as exemplified by Xanthomonas campestris and Burkholderia spp., and the omnipresent probiotic Lactobacilli, as well as environmentally important photoautotrophic cyanobacteria, the multicellular Myxococcus xanthus, and chemolithotrophic Acidithiobacillus are among the representatives of the microbial kingdom that are described. In turn, the various aspects of bacterial physiology affected by these signaling systems- e.g. biofilm formation and dispersal, the cell cycle, motility, virulence, production of antimicrobials, fundamental metabolism and osmohomeostasis - are discussed in detail in the context of different microorganisms. Dedicated chapters focus on the population diversity of cyclic dinucleotide signaling systems, their tendency to be horizontally transferred, the cyclic di-GMP signaling system in the social amoeba Dictyostelium, honorary cyclic (di)nucleotides, and the development of strategies for interfering with cyclic dinucleotide signaling in order to manipulate microbial behavior. Taken together, the chapters provide an authoritative source of information for a broad readership: beginners and advanced researchers from various disciplines; individuals seeking a broad overview of cyclic di-nucleotide signaling; and those who want to learn more about specific aspects. Also featuring reviews with a forward-looking perspective, the book offers a valuable source of inspiration for future research directions.

This book highlights key technologies and identifies areas for further development in proteogenomics. The utility and usefulness of very large Omics data sets (Next Gen Sequencing of DNA, RNA-seq, ribosome profiling, mass spectrometry- and antibody-based proteomics) is discussed and opportunities and challenges of related bioinformatics applications are outlined. The reader will be able to appreciate the interdisciplinary nature of the continuously evolving area of proteogenomics, which has already grown beyond its original concept of verifying gene annotations by proteomics. The chapters presented in this book are arranged to offer a general overview, rather than to provide detailed descriptions of technologies. The selected applications will provide useful insight into the level of detail that can be obtained in relation to certain diseases areas, including cancer biology and personalized medicine. The readers will find that each chapter delivers a comprehensive approach to proteogenomics, each from the point of view of a specific application. Research scientists interested in innovative processes that can offer a unique and at the same time a more complete access to technological developments and concepts that in turn can contribute to a better understand biological functions should read this book.

This fully updated edition provides a series of methods for how best to assess functions of histone deacetylases and acetyltransferases. The disease-relevance of dysregulated protein deacetylation by overexpressed or aberrantly activated histone deacetylases has spurred an intense search for novel and improved inhibitors of these enzymes, as reflected in this collection. Expert contributors explore the generation and evaluation of novel histone deacetylase inhibitors and new and improved techniques to assess acetylation-dependent molecular mechanisms in vitro and in vivo. 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, HDAC/HAT Function Assessment and Inhibitor Development: Methods and Protocols, Second Edition serves as an ideal guide for researchers seeking to further elucidate this vital area of study.

The incentive for putting together Volume 4 of this series was to review the wealth of new information that has become available in prokaryotic organisms in protein export and membrane biogenesis. Just in the last several years, protein translocation has now been efficiently reconstituted using defined components and the mechanism by which proteins are moved across membrane bilayers is now being examined at a higher resolution. In addition, because of a new technical breakthrough using osmolytes, it is now possible to reconstitute a number of channel proteins, ATPase, receptors, and transporters. In many cases, it is possible to successfully predict the membrane topology of these types of proteins using both "hydrophobicity analysis" and the "positive inside" rule.
In this volume, two chapters focus on protein translocation across membranes ("Biochemical Analyses of Components Comprising the Protein Translocation Machinery of E. Coli; Protein Translocation Genetics"), while several others on how proteins assemble into the ineer membrane of E. Coli ("Membrane Protein Assembly; Membrane Insertion of Small Proteins: Evolutionary and Functional Aspects; Pigment-Protein Complex Assembly in Rhodobacter sphaeroides and Rhodobacter Capsulatus"). Other sections review recent progress on transporters ("Identification and Reconstitution of Anion Exchange Mechanisms in Bacteria; Helic Packing in the C-Terminal Half of Lactose Permease") and signal transduction ("Mechanism of Transmembrane Signaling in Osmoregulation") as well as the assembly of prints into the outer membrane ("Export and Assembly of Outer Membrane Proteins in E. coli"). Although the emphasis of the book is on proteins, the role of phospholipids in controlling various cell surface processes is reviewed ("Role of Phospholipids in coli Cell Function"). I should point out the reason for the rapid progress in bacteria research is because of the possibility to apply biochemistry and genetics in this organism.

Proteins are the functional units of the cellular machinery and they provide significant information regarding the molecular basis of health and disease. Therefore, techniques to separate and isolate the various proteins are critical to studying and understanding their functional characteristics. One of the widely used techniques for this purpose is electrophoresis. In Protein Electrophoresis: Methods and Protocols, contributions from experts in the field have been collected in order to provide practical guidelines to this complex study. Each chapter outlines a specific electrophoretic variant in detail so that laboratory scientists may perform a technique new to their lab without difficulty. Written in the successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Protein Electrophoresis: Methods and Protocols seeks to serve laboratory scientists with well-honed, detailed methodologies in an effort to further our knowledge of this essential field.

In "Single Molecule Studies of Proteins," expert researchers discuss the successful application of single-molecule techniques to a wide range of biological events, such as the imaging and mapping of cell surface receptors, the analysis of the unfolding and folding pathways of single proteins, the analysis interaction forces between biomolecules, the study of enzyme catalysis or the visualization of molecular motors in action. The chapters are aimed at established investigators and post-doctoral researchers in the life sciences wanting to pursue research in the various areas in which single-molecule approaches are important; this volume also remains accessible to advanced graduate students seeking similar research goals.

Protein Glycosylation provides clear, up-to-date, and integrated coverage of key topics in this field. Particular emphasis is placed on the biosynthetic pathways that result in a wide variety of identified protein-bound oligosaccharides. Protein Glycosylation begins with an overview of the chemical structures of mono- and oligosaccharides, to provide a scientific basis for the later chapters. The book includes discussions on the purification, function, and enzyme kinetics of selected glycosidases and glycotransferases, as well as a review of the roles of oligosaccharides in glycoprotein function and the in vivo role of glycoproteins themselves. Finally, the in vitro synthesis of glycoproteins is presented, together with future directions in glycobiology. Protein Glycosylation serves as an excellent text for upper-level undergraduate and graduate students as well as a reference for those scientists whose training is not in glycobiology but who are moving into this field.

After the identification of a potential protein drug, the next critical step is the production of sufficient authentic material for testing, characterization, and clinical trials, which, when successful, leads to the need for robust methodologies for large-scale production, purification, characterization, viral inactivation, and continued testing of the final protein product. Building on the valuable first edition, Therapeutic Proteins: Methods and Protocols, Second Edition aims to cover each of these key aspects of protein drug production through the contributions of authors from highly esteemed industrial and academic institutions around the world. Emphasizing the newest developments in the field, this second edition also includes additional emphasis on discovery, including new display and screening methods as well as the design and engineering of new types of therapeutic proteins. There is also discussion of computational and bioinformatics methods, and chapters on safety aspects of therapeutic protein development. Written in the highly successful Methods in Molecular Biology (TM) format, protocol 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. Fully updated and practical, Therapeutic Proteins: Methods and Protocols, Second Edition provides an essential resource to all scientists working in the field of therapeutic proteins.

Protein misfolding is a key feature of many disorders in humans, given that over twenty proteins are known to misfold and cause disease. In "Protein Folding, Misfolding, and Disease: Methods and Protocols," experts in the field present a collection of current methods for studying the analysis of protein folding and misfolding, featuring strategies for expressing and refolding recombinant proteins which can then be utilized in subsequent experiments. This detailed volume also covers methods for analyzing the formation of amyloid, protocols for determining the size and structure of native and misfolded proteins, as well as specific examples of where misfolded proteins can be examined using state-of -the-art technologies. Written in the highly successful "Methods in Molecular Biology " series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Up to date and authoritative, "Protein Folding, Misfolding, and Disease: Methods and Protocols" offers researchers the tools necessary to move ahead in this vital field."

This volume covers a wide spectrum of techniques and approaches that are used in the upstream and downstream processing for recombinant glycoprotein production. Chapters guide the reader through state-of-art of therapeutic recombinant glycoproteins, explores the patent literature, expression systems used for glycoproteins production, methods employed in the downstream processing of different glycoproteins, and information about analytical tools and formulation strategies. 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. Authoritative and cutting-edge, Recombinant Glycoprotein Production: Methods and Protocols aims to ensure successful results in the further study of this vital field

This volume focuses on the structure, function and regulation of plant signaling G proteins and their function in hormonal pathways, polarity, differentiation, morphogenesis and responses to biotic and abiotic stresses.

Plants are sessile organisms that need to continuously coordinate between external and internal cues. This coordination requires the existence of hubs to allow cross-talk between different signaling pathways. A single family of Rho GTPases, termed either ROPS or RACs, and heterotrimeric G proteins have emerged as the major molecular switches in a multitude of signal transduction pathway in plants.

This book surveys the current knowledge concerning the expression and function of small stress proteins (sHsps) in different organisms, ranging from prokaryotes to humans. It provides an overview of the diversity and complex evolutionary history of sHsps and describes their function and expression in different eukaryote models. Additional chapters discuss the involvement of sHsps in pathological conditions and gene therapy approaches towards a control of sHsp expression levels.

Cancerremainstobeoneofthemostdevastatingdiseasesworldwidesincelong ago. Thepoorprognosisofcancerislargelyduetometastasis. Metastasisisoften depictedasamultistageprocessinwhichmalignantcellsspreadfromtheprimary locustodistantorgansviacirculation. Whereasgeneticalterationsweresuggested tobeessentialfortransformationofprimarytumorcellsintometastaticphenotype, epigeneticeventsareequallyimportant,whichmaybetriggeredbymetastaticf- torswhereverintheprimarytumorlocus,bloodcirculationandthesecondaryloci. Signaltransductionsinitiatedbythemetastaticfactorsareresponsibleformediating themolecularandcellularprocessesleadingtometastasis. Blockadeoftherelevant molecularpathwaysisoneofthemosteffectivestrategiesforpreventionoftumor metastasis. Clinicaltrialsareunderwaywithpromisingoutcome. Inthisbook, wetakecomprehensive review inregard withthisexciting eld ofcancerresearch. Chapter1takesabriefoverviewofrecentlyidenti edsignal mechanismsforeachstepoftumormetastasisincludingtheinitiationstage,intra- sation,anti-anoikisinbloodcirculation,homing,extravasationand nalsurvivalin themetastaticsite. Chapter2makesacompletedreviewforthemolecularandcel- lareventsinvolvedininitiationofmetastasis. Especially,thesignalingmechanisms formediatingtumorprogressioninducedbysomeimportantmetastaticfactorsare described. InChapters3and4,thecentralrolesofMAPKanditsdownstreameff- torsMAPKAPKplayineachstepoftumormetastasisarewelldelineated. Chapter5 furtherdescribesdetailedlyabouthowGrb2andotheradaptorproteins,upstreamof MAPK cascade, contribute tometastasis. InChapter 6, therole ofreactive o- genspecies(ROS)intumorprogressionarehighlighted. Moreover,thepotential contribution of ROS to cross talk between major signaling cascades that lead to sustainedMAPKactivationareproposedinChapter7. Chapter8takesaninsight intothesignalingmechanismsfordynamictraf ckingandturnoveroffocalad- sionproteinsinregulationoftractionandretractionforces,whichareneededfor celllocomotionandinvasion. Chapter9describestheinvolvementofNotchsign- ingpathwaywhichisnotonlyessentialforembryonicdevelopmentbutalsoplays importantroleintumorprogression. Chapter10reviewedtherecentlyidenti ed cancer- and metastasis-initiating cells involved in tumor progression. Especially, signal pathways that are frequently deregulated in cancer stem/progenitor cells v vi Preface duringcancerprogressionarehighlighted. Chapter11describestheroleoflipid rafts, a special component within membrane lipid domain, in signal transd- tion triggered by growth factor receptors leading to tumor metastasis. Finally, Chapters12,Chapters13,andChapters14presentthesignalingpathwaysresp- sibleformetastaticprogressionofspeci ctumorsincludingovariancancer,uveal melanomaandhepatoma,respectively. WethankallthecontributorsofeveryChapterinthebookincludingJia-RuWu, Chi-TanHu,LaureVoisin,StephanieDuhamel,SylvainMeloche,AlexeyShiryaev, MarijkeVanGhelue,UgoMoens,AlessioGiubellino,PraveenR. Arany,Moulay A. Alaoui-Jamali, Krikor Bijian, Panagiota Toliopoulos, Pingyu Zhang, Patrick A. Zweidler-McKay,MurielleMimeault,SurinderK. Batra,SamirKumarPatra, LydiaW. T. Cheung,CarmanK. M. Ip,AliceS. T. Wong,CecileLaurent,Jerome Couturier,XavierSastre-Garau,LaurenceDesjardins,EmmanuelBarillot,Sophie Piperno-Neumann,SimonSauleandRajagopalN. Aravalli. Wehopethisbookmightstimulatemorecancerbiologiststoemphasizethis eld whichbene tsdevisingmoreeffectivemoleculartargetingstrategiesforprevention ofcancermetastasis. Hualien,Taiwan Wen-ShengWu Chi-TanHu Contents 1 Overview of Signal Transduction in Tumor Metastasis...1 Wen-ShengWuandJia-RuWu 2 Microenvironment Triggers EMT, Migration and Invasion of Primary Tumor via Multiple Signal Pathways ...9 Wen-ShengWuandChi-TanHu 3 The ERK1/2 MAP Kinase Signaling Pathway in Tumor Progression and Metastasis ...25 LaureVoisin,StephanieDuhamel,andSylvainMeloche 4 Mitogen-Activated Protein Kinase-Activated Protein Kinases and Metastasis...41 AlexeyShiryaev,MarijkeVanGhelue,andUgoMoens 5 Grb2 and Other Adaptor Proteins in Tumor Metastasis ...77 AlessioGiubellinoandPraveenR. Arany 6 The Role of ROS Signaling in Tumor Progression...103 Wen-ShengWuandJia-RuWu 7 Signal Cross Talks for Sustained MAPK Activation and Cell Migration Mediated by Reactive Oxygen Species: The Involvement in Tumor Progression...

The biological interactions of living organisms, and protein-protein interactions in particular, are astonishingly diverse and present numerous challenges to modern biomolecular research because of their complexity. Analysis of patterns and principles governing these interactions has prompted a rapid development of computational methods to identify protein interaction partners and to understand the roles of individual components of protein interaction networks in cell functions.

This book integrates different approaches from bioinformatics, biochemistry, computational analysis and systems biology to offer the reader a comprehensive global view of the diverse data on protein-protein interactions and protein interaction networks. It brings together the descriptions of experimental techniques and expounds on different computational algorithms for protein network analysis and prediction of protein and domain interactions, with each chapter containing a description of the problem, a review of methods and algorithms, a list of resources and current conclusions.

Features

a [ Reviews experimental techniques for identification of protein interactions

a [ Discusses protein interaction databases and methods of integrating data from diverse sources

a [ Describes computational methods to predict protein and domain interaction partners

a [ Explores the properties of interaction interfaces and highlights approaches to model the assembly of protein complexes

a [ Examines the topological and dynamical properties of protein interaction networks and presents the tools for comparative analysis of these networks

Written by leading experts, Protein-protein Interactions and Networksprovides a broad, thorough and multidisciplinary coverage of this field. It will be invaluable to researchers from academia and the bioinformatics industry, as well as an excellent auxiliary text for graduate students studying the topic.

Knowledge of cholesterol and its interaction with protein molecules is of fundamental importance in both animal and human biology. This book contains 22 chapters, dealing in depth with structural and functional aspects of the currently known and extremely diverse unrelated families of cholesterol-binding and cholesterol transport proteins. By drawing together this range of topics the Editor has attempted to correlate this broad field of study for the first time. Technical aspects are given considerable emphasis, particularly in relation cholesterol reporter molecules and to the isolation and study of membrane cholesterol- and sphingomyelin-rich "raft" domains. Cell biological, biochemical and clinical topics are included in this book, which serve to emphasize the acknowledged and important benefits to be gained from the study of cholesterol and cholesterol-binding proteins within the biomedical sciences and the involvement of cholesterol in several clinical disorders. It is hoped that by presenting this topic in this integrated manner that an appreciation of the fact that there is much more that needs to be taken into account, studied and understood than the widely discussed "bad and good cholesterol" associated, respectively, with the low- and high-density lipoproteins, LDL and HDL. Content Level Professional/practitioner

The proteolytic enzymes have an essential function in all cells. Their activities are regulated by the rate of synthesis, activation of proenzymes and by the rate of synthesis of their inhibitors. They are synthesized in ribosomes like any other proteins and transported to various storage organelles or secreted from the cells and are activated in the pericellular space or in interstitium. Various cells and tissues have their characteristic enzyme patterns which serve their specific functions. Proteolytic enzymes take part and often have a regulatory role in numerous phases of cell function, e.g. cell division, migration, apoptotic as well as necrotic cell death etc. Diseases in which proteolysis has been subject of active research are e.g. cancer metastasis, viral infections, e.g. HIV, and Alzheimer's disease. They are also an essential part in any tissue remodelling, wound healing, throughout the kingdom of fauna and flora.

The aim of the Protein Reviews is to serve as a publication vehicle for review articles that focus on crucial current vigorous aspects of protein structure, function, evolution and genetics. The volumes will appear online before they are published in a printed book. Articles are selected according to their importance to the understanding of biological systems, their relevance to the unravelling of issues associated with health and disease or their impact on scientific or technological advances and developments. Volume 19 focusses on Purinergic receptors, also termed purinoceptors. These are plasma membrane proteins present in nearly all mammalian tissues. They participate in a number of cell functions that include proliferation and migration of neural stem cells, vascular reactivity, apoptosis and cytokine secretion and have been associated with learning and memory, feeding conduct, movement and sleep. They facilitate relaxation of smooth muscle of the gut in response to adenosine (P1 receptors) or ATP (P2 receptors). The chapters in this volume are authored by experts in the field. They deal with aspects of structure and biological activity of selected receptor proteins. The first chapter in this volume reviews the current research on the Mechanism of channel gating and regulation of the activity of calcium-activated chloride channel ANO1. This is followed by a chapter dealing with Structure and function of the two-component cytotoxins of Staphylococcus aureus and a chapter on Membrane Fusion and Infection involving the Influenza virus Hemagglutinin. The fourth chapter reviews the impact of arrhythmogenic mutations through the structural determination of the L-type voltage-gated calcium channel. Then there is a chapter that discusses some open questions pertaining to histone post-translational modifications and nucleosome organization in transcriptional regulation. The next chapter deals with regulation of the extracellular SERPINA5 (protein C inhibitor) penetration through cellular membranes. This is followed by a chapter on coding of Class I and II aminoacyl-tRNA synthetases; a chapter on regulation of nephrin phosphorylation in diabetes and chronic kidney injury and a chapter on The Structure-Forming Juncture in oxidative protein folding and the events in the ER. Finally the last chapter deals with the polyspecificity of anti-lipid antibodies and its relevance to the development of autoimmunity. This volume is intended for research scientists, clinicians, physicians and graduate students in the fields of biochemistry, cell biology, molecular biology, immunology and genetics.

The purpose of Calpain Methods and Protocols is quite straightf- ward: it is to present the actual experimental methods used in many different laboratories for the study of calpain. It will provide the vital experimental detail, and the discussion of possible pitfalls, for which the standard journals no longer provide space. This will make it as easy as possible for investi- tors interested in calpain to adopt established methods without repeating old mistakes, and to adapt and apply these methods in novel approaches to the many outstanding calpain questions. These questions range from purely biochemical problems of protein structure and enzyme regulation at the molecular level, through large areas of cell biology, to applied and clinical aspects of calpain function in human d- ease. Within this panoply of topics, a wide range of investigators will find many fascinating and as yet unanswered questions about calpain. Calpain Methods and Protocols will provide instant access to many essential te- niques, while saving them the time and effort involved in developing a new method. In addition to questions relating to the normal physiological roles of the calpains, there is considerable evidence that inappropriate calpain activity may have pathological effects in many tissues, for example, following ischemia. This provides a major stimulus for the development of specific calpain inhi- tors for therapeutic purposes, and for the development of methods to evaluate such inhibitors.

This edited book provides the first comprehensive overview on conventional and emerging processing technologies for the extraction and purification of proteins and/or peptides from plant sources with a special focus on subsequent product development. The book opens with an introduction to the most conventional processing technologies used in industry today: the alkaline extraction followed by isoelectric precipitation, and air classification. The book also focusses on novel extraction and purification technologies, covering the most recent green emerging technologies based on enzymatic processes, solvents, high-pressure processing, barometric membrane technologies, and microwave-assisted extraction, among others. The final chapters bridge the gap between the presented methods and product development and highlight how these technologies can alter protein functionality and nutritional quality of the extracted protein, and thereby, impact human health. In the context of rising consumer interest in foods from plant-protein ingredients and the United Nations targets for Sustainable Development Goal 12 on 'Responsible Consumption and Production', this book will provide an indispensable resource for students, engineers and researchers in academia and industry, working in the area of food science, food technology and plant-based product development.