This volume compiles new experimental approaches and concepts focusing mostly, but not solely, on ways to manipulate and regulate Ras activity and its downstream signaling output. Chapters detail standard methodologies, biochemical methods, Ras processing trafficking and localization, Ras signaling and inhibition, and in vivo models for studying Ras function. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Ras Activity and Signaling: Methods and Protocols aims to provide support and guidance to lab workers in their work on Ras GTPases and in the design of new projects requiring novel methodologies.

This book reviews the latest trends in glycobiotechnology, it offers an authoritative discussion about future directions of glycoengineering, and it provides a comprehensive overview about the current and emerging approaches to identify, quantify and characterize glycosylated proteins. Divided into 14 chapters, the book outlines recombinant glycoprotein expression in mammalian cells, insect cells, yeast, and bacterial systems. It covers the chemical and enzymatic syntheses of glycans and glyconjugates, and addresses the impact of glycosylation on protein function for the development of biologicals including vaccines. In the final chapters of the book, readers will discover more about the state-of-the-art in glycomics, glycoproteomics and glycan array technologies.

The origin of life has been investigated by many researchers from various research fields, such as Geology, Geochemistry, Physics, Chemistry, Molecular Biology, Astronomy and so on. Nevertheless, the origin of life remains unsolved. One of the reasons for this could be attributed to the different approaches that researchers have used to understand the events that happened on the primitive Earth. The origins of the main three members of the fundamental life system, as gene, genetic code and protein, could be only separately understood with these approaches. Therefore, it is necessary to understand the origins of gene, the genetic code, tRNA, metabolism, cell structure and protein not separately but comprehensively under a common concept in order to understand the origin of life, because the six members are intimately related to each other. In this monograph, the author offers a comprehensive hypothesis to explain the origin of life under a common concept. At the same time, the author offers the [GADV] hypothesis contrasting it with other current hypotheses and discusses the results of analyses of genes/proteins and the experimental data available in the exploration of the current knowledge in the field. This book is of interest for science students, researchers and the general public interested in the origin of life.

Leading researchers are specially invited to provide a complete understanding of a key topic within the multidisciplinary fields of physiology, biochemistry and pharmacology. In a form immediately useful to scientists, this periodical aims to filter, highlight and review the latest developments in these rapidly advancing fields. Chapter "Stationary and Non-Stationary Ion- and Water Flux Interactions in Kidney Proximal Tubule. Mathematical Analysis of Isosmotic Transport by a Minimalistic Model" is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Medicinal chemistry is both science and art. The science of medicinal chemistry offers mankind one of its best hopes for improving the quality of life. The art of medicinal chemistry continues to challenge its practitioners with the need for both intuition and experience to discover new drugs. Hence sharing the experience of drug research is uniquely beneficial to the field of medicinal chemistry. Drug research requires interdisciplinary team-work at the interface between chemistry, biology and medicine. Therefore, the topic-related series Topics in Medicinal Chemistry covers all relevant aspects of drug research, e.g. pathobiochemistry of diseases, identification and validation of (emerging) drug targets, structural biology, drugability of targets, drug design approaches, chemogenomics, synthetic chemistry including combinatorial methods, bioorganic chemistry, natural compounds, high-throughput screening, pharmacological in vitro and in vivo investigations, drug-receptor interactions on the molecular level, structure-activity relationships, drug absorption, distribution, metabolism, elimination, toxicology and pharmacogenomics. In general, special volumes are edited by well known guest editors.

This volume provides a comprehensive set of protocols that can be used by any research lab to investigate diverse functional and structural properties of Single Stranded DNA Binding Proteins (SSBs) from eubacterial, archaeal, eukaryotic, mitochondrial and viral systems. 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, Single Stranded DNA Binding Proteins aims to be a useful practical guide to researchers to help further their study in this field.

This work is concerned with a group of proteins which were originally consid ered to be an esoteric phenomenon but which have now been shown to play critical roles both in normal and stressed cells as well as being involved in a variety of human diseases. It is the purpose of this work to give a comprehen sive view of these proteins and their various aspects. After an introductory chapter providing an overview of these proteins, the work is divided into four main sections each of which deals with one important aspect of these proteins. Thus, the first section contains a series of chapters which describe individual stress proteins and their roles in particular biological phenomena. Evidently, the induction of these proteins by elevated tempera ture or other stresses is their defining feature and the second section of this book therefore considers the regulation of stress protein gene expression both by stressful stimuli such as elevated temperature or ischaemia and by non stressful stimuli such as cytokines.

The book Heat Shock Proteins in Cancer Therapeutics provides the most comprehensive review on contemporary knowledge on the role of HSP in various types of cancer therapeutics. Using an integrative approach, the contributors provide a synopsis of the most current updates on the state of HSP in cancer therapeutics. The heat shock response pathway is a highly conserved cellular process. Heat shock factors are a master transcriptional regulator responsible for expression of several important heat shock proteins, which can effectively protect critical client proteins from misfolding and degradation, thus maintaining intracellular integrity under stressed conditions. Recent studies have demonstrated the direct connections between heat shock response players and tumor cell survival, validating heat shock response players as novel molecular targets in anticancer treatment. Although many hurdles in clinical application still need to be effectively addressed, such as undesirable drug toxicity and off target effects; narrow therapeutic window; poor PK/PD profiles, etc. Recent reports on synergistic drug combination, advanced prodrug design, smart nanoparticle packaging, and RNA aptamer selection offer promising solutions to overcome these challenges. Future advancements in this fast-growing area can potentially lead to the next generation of cancer therapeutics. Key basic and clinical research laboratories from major universities, academic medical hospitals, biotechnology and pharmaceutical laboratories around the world have contributed chapters that review present research activity and importantly project the field into the future. The book is a must read for graduate students. medical students, basic science researchers and postdoctoral scholars in the fields of Cancer Biology, Oncology, Translational Medicine, Clinical Research, Biotechnology, Cell & Molecular Medicine, Pharmaceutical Scientists and Researchers involved in Drug Discovery.

In this thesis, the author outlines the development of new monolithic columns and isotope dimethyl labeling strategies and their applications in high-performance proteome analyses. Though different types of monolithic columns have been widely developed for chromatography and electrophoresis separation, their application in proteomics for complex peptide mixtures separation is still a challenge. The author discusses the preparation of new monolithic columns and optimization of chromatography separation capability to improve coverage and accuracy of proteome analysis. Further, the author describes a novel online multidimensional chromatography system combined with automated online isotope labeling, which significantly improves the throughput, sensitivity and accuracy of quantitative proteomics. In addition to the development of new technologies, the author investigates the proteome and phosphoproteome expression changes of clinical hepatocellular carcinoma tissues and the hippocampi of mice with Alzheimer s disease. The work in this thesis has led to several publications in high-profile journals in the fields of analytical chemistry and proteome research."

This book fills the need for a simplified text covering western blotting protocols aimed not just at high school and college students, but the researcher with little to no experience in these techniques. It provides the principles, basic methodology, and tips and tricks to avoiding the common pitfalls of western blotting. The book also introduces simple protocols that can transform western blotting into a fun method, such as sending secret messages on membranes or using nitrocellulose membrane as a canvas for art. In addition to the techniques, this book also covers the history of western blotting, which originated from the development of the blotting of DNA. It then delves into the importance of protein blotting, brought to the fore by the fact that the procedure has been evolving constantly since its inception in 1979, and the fact that the scientific community is faced with a multitude of ways and means of transferring proteins to membranes..

This book addresses the differentiation control of skeletal muscle in different locations of the vertebrate body Particular attention is paid to novel regulatory molecules and signals as well as the heterogeneity of origin that have revealed a developmental overlap between skeletal and cardiac muscle. Different functional muscle groups are the product of the evolution of the vertebrate classes, making a phylogenetic comparison worthwhile for understanding the role of muscle stem cells and precursors in myogenesis. New insights into the hierarchy of transcription factors, particularly in the context of these different muscle groups have been gained from detailed investigations of the spatio-temporal and regulatory relationships derived from mouse and zebrafish genetics and avian microsurgery. Importantly, epigenetic mechanisms that have surfaced recently, in particular the role of MyomiRs, are also surveyed. With an eye to the human patient, encouraging results have been generated that identify parallels between embryonic myogenesis and regenerating myofibers due to common regulatory molecules. On the other hand, both processes differ considerably in quality and complexity of the processes employed. Interestingly, the heterogeneity in embryonic sources from which skeletal muscle groups in the vertebrate including the human body take origin is paralleled by differences in their susceptibility to particular muscle dystrophies as well as by the characteristics of the satellite cells involved in regeneration. The progress that has been made in the field of muscle stem cell biology, with special focus on the satellite cells, is outlined in this book by experts in the field. The authors review recent insights of the heterogeneous nature of these satellite cells regarding their gene signatures and regeneration potential. Furthermore, an improved understanding of muscle stem cells seems only possible when we study the impact of the cell environment on efficient stem cell replacement therapies for muscular dystrophies, putting embryological findings from different vertebrate classes and stem cell approaches into context.

Signal transduction is the fundamental mechanism for regulation of cellular activities by environmental cues and regulatory signals, and is particularly important for plants, whose survival requires proper physiological and developmental responses to the environmental changes. Much progress has been made recently in the plant signal transduction research field thanks to the development of diverse techniques which are covered in Plant Signalling Networks: Methods and Protocols. These include advanced research methods such as proteomics and mass spectrometry methods for studying protein modification, biochemical and cell biological tools for studying protein-protein interactions, genomic techniques for dissecting protein-DNA interaction and transcription networks, and computation methods that integrate molecular network into plant developmental processes. 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. Plant Signalling Networks: Methods and Protocols presents well-honed methodologies for a wide range of research approaches including genetics, proteomics, biochemical, cell biological, and computational approaches, and seeks to serve both professionals and novices with a comprehensive understanding of complex signaling networks in plants.

A broad definition of a receptor is a specialized protein on or in a cell that recognizes and binds a specific ligand to undergo a conformational change, leading to a physiological response or change in cell function. A ligand can be an endogenous neurotransmitter, hormone, paracrine/autocrine factor, or a synthetic drug that may function as an agonist or antagonist. The third edition of Receptor Binding Techniques expands upon the methods and techniques used for studying receptors in silico, in vitro and in vivo. Comprehensive chapters describe how to use online resources for experimental research such as prediction of receptor-ligand interactions and mine the IUPHAR receptor database. Classical techniques of radioligand binding, quantitative autoradiography and their analyses are complemented by the use of immunocytochemistry for the cellular localization of receptor protein and hybridization to detect receptor mRNA. Protocols using fluorescent labeled ligands are described to visualise receptors in living cells, their interaction with beta-arrestin to measure ligand-induced internalisation and green fluorescent protein to study trafficking. Non-radioactive, chemiluminescent cAMP and arrestin assays facilitate the identification of novel 'biased agonists'. Detailed methods are provided for in vivo imaging of receptors using positron emission tomography (PET). Written in the highly 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 laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Receptor Binding Techniques, Third Edition, aids scientists in continuing to study receptor binding.

This detailed new edition presents the latest developments of the main pillars of protein analysis, namely sample preparation, separation, and characterization. Core areas in this volume are protocols for the analysis of post-translational modifications and protein interaction partners, followed by sophisticated procedures to enrich for extracellular vesicles and organelles, along with several types of protein immuno-assays complemented by various methods for the characterization of antibodies and host-cell protein analysis. Last but not least, a few standard sample preparation protocols and recent advances concerning immuno-chemical detection of proteins are included as well. 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 up-to-date, Proteomic Profiling: Methods and Protocols, Second Edition serves as an ideal reference for students of biochemistry, biomedicine, biology, and genomics and will be an invaluable source for the experienced, practicing scientist as well.

This detailed volume explores fibrous proteins widely present in different biological tissues or biological structural materials. The book begins by introducing the structure of representative fibrous proteins, including animal silks, collagen, elastin, resilin, and keratin, and it then continues by providing detailed experimental protocols for the synthesis, assembly, and characterization of natural, regenerated, and recombinant fibrous proteins. 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, Fibrous Proteins: Design, Synthesis, and Assembly is an ideal guide for researchers aiming to master fibrous protein preparations with the aid of this broad and interdisciplinary perspective on understanding the structure-property-function relationships of natural and reconstituted fibrous proteins.

In this book, the major paradigm-shifting discoveries made in the past century on key cellular nanomachines are described in great detail: their complex yet precise and elegant design and function, as well as the diseases linked to their dysfunction and the therapeutic approaches to overcome them. The major focus of this book is the "porosome" nanomachine, the universal secretory portal in cells. This is an ideal book for students, researchers, and professionals in the fields of nanoscience and nanotechnology.

Protein Phosphatase Protocols presents a broad range of protocols for the study of protein phosphatases, all written by experts and innovators from phosphatase laboratories around the world. This volume is a compendium of resources for the study of protein phosphatases and their potential as drug targets. Experimental methodologies are taken from proteomics, bioinformatics, genomics, biochemistry, RNAi, and genetics.

This book reviews the principles of design and examples of successful implementation of proteinkinase inhibitors (PKI), and offers a comprehensive and authoritative overview of the history and latest developments in the field. Chapters written by experts from industry and academia cover the function, structure and topology of Proteinkinases, molecular modelling, disclose how to achieve high level of selectivity for kinase inhibitors, and exploit kinase inhibitors for cancer treatment. Particular attention is given to Inhibitors of c-Jun N-terminal kinase 3, and to covalent Janus Kinase 3 Inhibitors. A case study on Receptor Tyrosine Kinases EGFR, VEGFR, PDGFR is also presented in this book. Given its breath, this book will appeal to medicinal chemists, students, researchers and professionals alike.

Recent advances in the field of peptide chemistry and gene technology have resulted in an explosive accumulation of information on biologically active pep tides and functional proteins. Because of the importance of such peptides and proteins in the role of cellular or extracellular regulatory mechanisms and their potential therapeutic value, an understanding of their detailed interactions with the specific receptor should provide useful information for structure-activity studies. These problems have been approached in many ways. However, despite our efforts, many gaps in our knowledge of peptide chemistry remain to be filled, and some answers will no doubt be forthcoming in the next few years. This volume, the Proceedings of the 2nd Japan Symposium on Peptide Chemistry held in Shizuoka, covers all presentations. Speakers and discussants, numbering approximately 550, came from Australia, Austria, Belgium, Canada, China, Denmark, France, Germany, Israel, Italy, Russia, Sweden, Switzerland, India, the United Kingdom, the United States of America, and Japan. One very sad note was the sudden death, shortly before the conference, of Professor Emeritus Shiro Akabori, an outstanding organic chemist and a pioneer in peptide research. The news shocked his many friends and colleagues, who miss him deeply. Finally, it is a pleasure to acknowledge the help of those individuals and organizations who made the conference possible: the contributing scientists; the advisory committee and the staff of the conference; the Japanese Peptide Society, and other institutions; and the corporations which gave their financial support.

This book summarizes the latest studies on plant reproduction and multiple aspects of signaling in reproductive development. It also presents the most advanced processes in CrRLK1L receptor and RALF peptide studies during plant development. Focusing on signaling in pollen tube integrity and sperm release regulation, it provides significant insights into the BUPS-ANX receptor complex and the corresponding ligands RALF4/19 to promote pollen tube growth with proper cell integrity. It also proposes a working model of female tissue-derived RALF34 competing with RALF4/19 from the BUPS-ANX to trigger pollen tube rupture and sperm release. Offering a detailed overview of the spatiotemporal regulation mechanism underlying the control of pollen tube integrity and sperm release, the book fills a major gap in our understanding of plant reproductive processes, and as such is a valuable resource for those working in the area of plant signaling.

This thorough volume explores predicting one-dimensional functional properties, functional sites in particular, from protein sequences, an area which is getting more and more attention. Beginning with secondary structure prediction based on sequence only, the book continues by exploring secondary structure prediction based on evolution information, prediction of solvent accessible surface areas and backbone torsion angles, model building, global structural properties, functional properties, as well as visualizing interior and protruding regions in proteins. Written for the highly successful Methods in Molecular Biology series, the chapters include the kind of detail and implementation advice to ensure success in the laboratory. Practical and authoritative, Prediction of Protein Secondary Structure serves as a vital guide to numerous state-of-the-art techniques that are useful for computational and experimental biologists.

The reproduction and spread of a virus during an epidemic proceeds when the virus attaches to a host cell and viral genetic material (VGM) (protein, DNA, RNA) enters the cell, then replicates, and perhaps mutates, in the cell. The movement of the VGM across the host cell outer membrane and within the host cell is a spatiotemporal dynamic process that is modeled in this book as a system of ordinary and partial differential equations (ODE/PDEs). The movement of the virus proteins through the cell membrane is modeled as a diffusion process expressed by the diffusion PDE (Fick's second law). Within the cell, the time variation of the VGM is modeled as ODEs. The evolution of the dependent variables is computed by the numerical integration of the ODE/PDEs starting from zero initial conditions (ICs). The departure of the dependent variables from zero is in response to the virus protein concentration at the outer membrane surface (the point at which the virus binds to the host cell). The numerical integration of the ODE/PDEs is performed with routines coded (programmed) in R, a quality, open-source scientific computing system that is readily available from the Internet. Formal mathematics is minimized, e.g., no theorems and proofs. Rather, the presentation is through detailed examples that the reader/researcher/analyst can execute on modest computers. The ODE/PDE dependent variables are displayed graphically with basic R plotting utilities. The R routines are available from a download link so that the example models can be executed without having to first study numerical methods and computer coding. The routines can then be applied to variations and extensions of the ODE/PDE model, such as changes in the parameters and the form of the model equations.

This edition details a collection of specific shotgun proteomics-based laboratory techniques and applications developed in leading laboratories and proteomics units worldwide. Chapters cover a broad range of topics covering, shotgun proteomics of extracellular vesicles and subcellular structures, shotgun proteomics in non-model organisms, clinical proteomics, food proteomics, analysis of post-translational modifications and protein complexes, and data processing and storage. 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, Shotgun Proteomics: Methods and Protocols aims to be an up-to-date guide for researchers seeking to understand the proteome of any given biological sample.

Christopher Schirwitz's thesis focuses on improving the quality of in situ synthesized high-complexity peptide micro arrays. Micro arrays containing proteins or small protein fragments in the form of peptides have become of great interest in proteomic research. With the help of these microarrays a large number of potential target molecules can be screened for interaction with a probe in a short timeframe. However, protein and peptide micro arrays are still lagging behind oligonucleotide arrays in terms of density, quality and manufacturing costs. A new approach developed at the German Cancer Research Center (DKFZ) has improved the synthesis of high-density peptide arrays. The current technology is capable of producing arrays with up to 40,000 different peptides per square cm by means of micro particle-based solid phase peptide synthesis. However, in situ synthesis approaches bear a conceptual disadvantage: The quality of the peptides is dependent on the efficiency of the synthesis so that peptide fragments are present in the resulting array among the desired full-length peptides. In peptide-protein interaction studies such peptide fragments. The central achievement of this thesis is the development of a new method allowing for the fast one-step purification of entire arrays without loss of resolution or spatial information. Christopher Schirwitz's work has resulted in a number of publications in high ranking journals.

This second edition presents an up-to-date chapters describing the most relevant and novel techniques employed to study the opioid receptors. Chapters detail transcriptional and post-transcriptional analysis, cellular detection of opioid receptors, analysis of signaling events modulated by opioid receptors, model systems to studying opioid receptor-mediated functions, and behavioral effects mediated by opioid receptors. 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, Opioid Receptors: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.