The Fifteen American Peptide Symposium (15APS) was held in Nashville, Tennessee, on June 14-19, 1997. This biennial meeting was jointly sponsored by the American Peptide Society and Vanderbilt University. The attendance of 1,081 participants from 37 countries was lower than the two previously held Symposia. However, the number of participating countries was the largest. Thus, it was gratifying to see that this meeting retained both its international flavor and participant loyalty at a time when there are many more symposia held each year on similar subjects. The scientific program, thanks to the insights and efforts of the Program Committee as well as Dr. Peter Schiller, the President of the American Peptide Society, was extraordinarily rich, diverse, and exciting. It was comprised of 124 oral and 550 poster presentations. Three prominent format changes were installed. First, the Symposium started on Saturday instead of Sunday. Second, the program opened on Saturday afternoon with a Mini-symposium by the Young Investigators to give them an early start and attention. Finally, 40 short and definitive reports were given in two parallel sessions. The expanded format permitted an unprecedented number of lectures and enabled wider participation by the attending delegates.

The first volume in this Methods Molecular Biology series, Proteins (1984), concentrated on basic techniques for the analysis and purification of peptides and proteins. As the series developed, more specialized volumes on proteins were introduced, such as those on Immunochemical Protocols (vol. 10), Practical Protein Chro- tography (vol. 11), Analysis Glycoprotein Biomedicine (vol. 14), Protein-DNA Interactions (vol. 30), Biomembrane Protocols (vols. 19 and 27), Analyses and Methods (vol. 17), and Optical Spectroscopy, Microscopy, and Macroscopic Techniques (vol. 22). Further specialist volumes on peptides, monoclonal antibodies, immunoassays, ELISA, protein engineering, protein stability, mass spectrometry of proteins, automated sequence analysis, and protein NMR are currently in preparation. Since it is now a decade since the initial volume was published, it seems an especially appropriate moment to extensively reorganize, update, and revise the earlier volume. In an attempt to be more c- prehensive in our coverage, this current volume, Basic Protein and Peptide Protocols, is totally committed to basic analytical methods; a planned companion volume will later concentrate on preparative techniques. Those analytical techniques requiring expensive speci- ized instrumentation, such as NMR, mass spectrometry, X-ray cr- tallography, spectroscopy, and automated sequence analysis, are not described here, but in the appropriate specialized volumes listed above.

This book brings together three decades worth of collaborative research to address the question "What sustains life?" In part a scientific response to Schroedinger's work "What is Life?" Urry's text contains elements of memoir, history, and a solid, informative scientific core that will interest both the general reader, student, and professional researcher.

This comprehensive, edited book explores carotenoids and their important functional roles in yeast, bacteria and plants and a profound exposition on the structures of carotenoid molecules, focusing in the first of three parts on the biosynthesis of carotenoids. The regulation of carotenoid biosynthesis in photosynthesis as well as in plant, fruits, storage roots and algae is central to the second part, and discoveries about the function of carotenoids in human health feature in the third and final part. Many helpful illustrations, explanations, overviews and examples help to bring readers up to date on relevant themes including carotenogenic genes, carotenoids in fruits and metabolic engineering. The book explores where carotenoids are synthesized in nature, including in carrots and algae. Contributing expert authors examine enzyme functions and plant models, and analyze the structure of carotenoid molecules. The function of carotenoids in photosynthesis and in photosynthetic organs as well as during fruit ripening are then explored. A whole chapter is dedicated to the latest research on apocarotenoids and further chapters cover interesting and novel themes on plastid development and the epigenetic regulation that affects carotenoid synthesis in plants. The metabolic engineering of carotenoids that has been done in fruits, plants, and seeds is another area that readers can explore, along with evidences on the function of carotenoids in human nutrition, as antioxidants, as in the control of lipid metabolism and in the absorption of carotenoids. This is a highly informative and wide-ranging work which will update researchers in the field, as well as supporting students of plant physiology and biotechnology, as supplementary reading.

Remarkably, while G protein-coupled receptors (GPCRs) are highly prevalent in animals and yeast, very few candidate GPCRs have been identified in plants. In G Protein-Coupled Receptor Signaling in Plants: Methods and Protocols, experts in the field describe techniques used in the study of small GTPases and related proteins. Beginning with a chapter on bioinformatics approaches for GPCR discovery, this detailed volume continues with chapters on heterotrimeric G protein subunits, Rab-GTPases, as well as lipid modifications, including myristoylation, acylation, and prenylation. 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. Practical and dependable, G Protein-Coupled Receptor Signaling in Plants: Methods and Protocols aims to aid further studies into the roles of small GTPases which will help elucidate numerous key processes in plants.

Recent advances in large scale DNA sequencing technology have made it possible to sequence the entire genome of an organism. Attention is now turning to the analysis of the product of the genome, the proteome, which is the set of proteins being expressed by a cell. Mass spectrometry is the method of choice for the rapid large-scale identification of these proteomes and their modifications. This is the first book to extensively cover the applications of mass spectrometry to proteome research.

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.

At the dawn of the 21st century, biotechnology is emerging as a key enabling technology for sustainable environmental protection and stewardship. Biotechnology for the Environment: Soil Remediation offers a state-of-the-art account of environmental biotechnology both in emerging and in more mature technological applications of soil remediation and cleanup of contaminated sites. Harnessing the potential of microorganisms and plants as eco-efficient and robust cleanup agents in a variety of practical situations is not only possible but is becoming widespread practice. Chapters are featured on current experience and trends in bioremediation of contaminated soil, life cycle assessment software tools for remediation planning, ex situ cleanup technologies using slurry reactors, implementation of anaerobic and aerobic in situ processes including monitored natural attenuation, complementary technologies on pesticide immobilisation in soil or humification of nitroaromatics, and, finally, phytoremediation of recalcitrant organic compounds and heavy metals. These contributions should be of value to environmental scientists, engineers and decision-makers involved in the development, evaluation or implementation of biological treatment systems.

For more information on Strategy and Fundamentals, see Focus on Biotechnology volume 3A, and for more information on Waste Water and Waste Gas Handling, see Focus on Biotechnology volume 3C.

This work presents a snapshot of the state of the art of modern biomolecular crystallography, from crystallisation through structure determination and even interactive presentation on the web. Methods driving the latest automated structure determination pipelines are explained, as well as how to deal with problems such as crystal pathologies that still demand expert analysis. These methods are illustrated through their application to problems of great biological interest, such as the molecular machinery underlying the complement pathway, the mechanism of action of monoamine oxidase inhibitors, and the structure of the eukaryotic ribosome. Complementary approaches, such as neutron diffraction, small angle X-ray scattering, coherent diffraction and computational modelling, are also explored.

Plant Proteomics: Methods and Protocols, Second Edition presents recent advances made in the field of proteomics and their application to plant biology and translational research. In recent years, improvements in techniques and protocols for high-throughput proteomics have been made at all workflow stages, from wet (sampling, tissue and cell fractionation, protein extraction, depletion, purification, separation, MS analysis, quantification) to dry lab (experimental design, algorithms for protein identification, bioinformatics tools for data analysis, databases, and repositories). Divided into nine convenient sections, chapters cover topics such as applications of gel-free, label- or label-free, imaging and targeted approaches to experimental model systems, crops and orphan species, as well as the study and analysis of PTMs, protein interactions, and specific families of proteins, and finally proteomics in translational research. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Plant Proteomics: Methods and Protocols, Second Edition seeks to serve both professionals and novices looking to exploit the full potential of proteomics in plant biology research.

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neurosclentlsts, and those that are available often seem either out of date or limited m scope. This series is about the methods most widely used by modern-day neurosclentists and 1s wrltten by their colleagues who are practicing experts. Volume 1 will be useful to all neurosclentists since It concerns those procedures used routinely across the widest range of subdisciplines. Collecting these general techniques together m a single volume strikes us not only as a service, but will no doubt prove of exceptIona utilitarian value as well. Volumes 2 and 3 - scribe all current procedures for the analyses of amines and their metabolltes and of ammo aads, respectively. These collections will clearly be of value to all neuroscientlsts working m or contemplating research m these fields. Similar reasons exist for Volume 4 on receptor binding techniques since experimental - tails are provided for all types of llgand-receptor bmdmg, including chapters on general prmciples, drug discovery and - velopment, and a most useful appendix on computer programs for Scatchard, nonlinear, and competitive displacement analyses. Volume 5 provides procedures for the assessment of enzymes - volved m biogenic amme synthesis and catabolism. Volumes in the NELJROMETHODS series will be useful to neurochemists, -pharmacologists, -physiologists, -anatomlsts, psychopharmacologists, psychlatnsts, neurologists, and chemists (organic, analytical, pharmaceutical, medicinal), in fact, everyone involved m the neurosclences, both basic and clinical.

Recent success stories demonstrated that the combination of subcellular prefractionation methods with proteomic analysis is a very potent approach to simplify complex protein extracts from cells or tissues and to detect low abundance proteins. It also made clear that sophisticated strategies encompassing sample preparation, analytics and validation steps were required to fully exploit the potential of subcellular proteomics.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.

Defining and understanding cellular and molecular mechanisms that are relevant to women's health has become a critical area of scientific pursuit. Until recently, very little effort has been place on defining or understanding critical differences between women and men that may be critical to the overall health of the woman. In 1990, the National Institutes of Health recognized this gap in knowledge resulting in the creation of the Office of Research on Women's Health. One of the purposes of this office was to advance the understanding of health issues from the women's perspective from both a basic and clinical scientific perspective. From a scientific evolution of understanding, the existence of this office is new and thus there has not been enough time for new information to integrate itself in our current scientific thought process. This book will seek to capture and disseminate our current understanding of scientific advancements relevant to women's health and provide the information to a broad audience. The purpose of this work is to discuss recent advancements in basic science across three areas of concern for women's health. In addition, the book will provide "translational" chapters that attempt to place the basic science work in context within our current understanding of the human. Although it is well acknowledge that gender differences exist across organ function which translates into differences in whole body function, until recently little effort has been made to define basic mechanisms within various tissues within the woman. This work will focus on recent scientific findings that are relevant to women's health and to provide novel and relevant information to interested scientists and clinicians.

Despite considerable variability within the scientific community, allosteric regulation can best be defined functionally as how a macromolecule binds one ligand differently when a second ligand is or is not pre-bound to the macromolecule, which constitutes a vital aspect of protein structure/function. In "Allostery: Methods and Protocols," expert researchers in the field provide key techniques to investigate this biological phenomenon. Focusing on heterotropic systems with some coverage of homotropic systems, this volume covers the monitoring of allosteric function, allosteric conformational changes, and allosteric changes in protein dynamics/sub-population distribution, as well as topics such as macromolecular and ligand engineering of allosteric functions and computational aids in the study of allostery. Written in the highly successful "Methods in Molecular Biology" series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory.

Thorough and intuitive, "Allostery: Methods and Protocols" aids scientists in continuing to study ligand-induced, through-protein effects on protein function (ligand binding/catalysis), a phenomenon that is well recognized through the history of the life sciences and very poorly understood at the molecular level."

This book describes more than 60 web-accessible computational tools for protein analysis and is totally practical, with detailed explanations on how to use these tools and interpret their results and minimal mentions to their theoretical basis (only when that is required for making a better use of them). It covers a wide range of tools for dealing with different aspects of proteins, from their sequences, to their three-dimensional structures, and the biological networks they are immersed in. The selection of tools is based on the experience of the authors that lead a protein bioinformatics facility in a large research centre, with the additional constraint that the tools should be accessible through standard web browsers without requiring the local installation of specific software, command-line tools, etc. The web tools covered include those aimed to retrieve protein information, look for similar proteins, generate pair-wise and multiple sequence alignments of protein sequences, work with protein domains and motifs, study the phylogeny of a family of proteins, retrieve, manipulate and visualize protein three-dimensional structures, predict protein structural features as well as whole three-dimensional structures, extract biological information from protein structures, summarize large protein sets, study protein interaction and metabolic networks, etc. The book is associated to a dynamic web site that will reflect changes in the web addresses of the tools, updates of these, etc. It also contains QR codes that can be scanned with any device to direct its browser to the tool web site. This monograph will be most valuable for researchers in experimental labs without specific knowledge on bioinformatics or computing.

Although the term redox covers an important number of chemical reactions, biochemists are more familiar with reactions involving the reactions mediated by electron transfer chains associated with respiration, the thiol-disulfide exchanges and the reactions occurring in the presence of free radicals. More recently, the importance of these reactions in the living world and in medicine has been recognized by biochemists, biologists, physiologists, physicians, etc. The importance of the subject in both fundamental and is reflected by the abundance of interesting reviews applied science concerning the subject (Cadenas, 1989, Del Maestro, 1991) and books (Dreosti, 1991; Rice-Evans and Burdon, 1994; Armstrong, 1994) The aim of this chapter is to describe basic reactions known with references to reviews covering special subjects related to redox reactions. Transformation of energy in living organisms is mediated by complex biological systems such as electron transfer chains where the succession of redox reactions provides energy to the organisms. Molecular oxygen or dioxygen is an essential molecule and is the terminal acceptor of electrons during respiration in eukaryotes. In these organisms, the electron transfer chain is located in the mitochondrial membranes and produces adenosine triphosphate (ATP). In anaerobes, the electron acceptor is C0 , S, sulphate or nitrate ions 2 instead of 02.

The genomes of cellular organisms are organized as double-stranded DNA, a structure that must be unwound to provide DNA replication, recombination, and repair machinery access to genomic information. However, DNA unwinding comes with inherent risks to genome stability. To help mediate these risks, bacterial, archael, and eukaryotic cells have evolved protective ssDNA-binding proteins (SSBs) that bind ssDNA with high affinity and specificity. SSBs also aid genome metabolic processes through direct interactions with key proteins in genome maintenance enzymes. Single-Stranded DNA Binding Proteins: Methods and Protocols assembles methods developed for examining the fundamental properties of SSBs and for exploiting the biochemical functions of SSBs for their use as in vitro and in vivo reagents. Clearly and concisely organized, the volume opens with an introduction to the structures and functions of SSBs, followed protocols for studying SSB/DNA complexes, methods for studying SSB/heterologous protein complexes, protocols for interrogating post-translational modifications of SSBs, and concludes with uses of fluorescently-labeled SSBs for in vitro and in vivo studies of genome maintenance 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. Authoritative and easily accessible, Single-Stranded DNA Binding Proteins: Methods and Protocols provides a rich introduction for investigators who are interested in this fascinating family of DNA-binding proteins.

When I was asked to edit the second edition of Protein NMR Techniques, my first thought was that the time was ripe for a new edition. The past several years have seen a surge in the development of novel methods that are truly revolutionizing our ability to characterize biological macromolecules in terms of speed, accuracy, and size limitations. I was particularly excited at the prospect of making these techniques accessible to all NMR labs and for the opportunity to ask the experts to divulge their hints and tips and to write, practically, about the methods. I commissioned 19 chapters with wide scope for Protein NMR Techniques, and the volume has been organized with numerous themes in mind. Chapters 1 and 2 deal with recombinant protein expression using two organisms, E. coli and P. pastoris, that can produce high yields of isotopically labeled protein at a reasonable cost. Staying with the idea of isotopic labeling, Chapter 3 describes methods for perdeuteration and site-specific protonation and is the first of several chapters in the book that is relevant to studies of higher molecular weight systems. A different, but equally powerful, method that uses molecular biology to "edit" the spectrum of a large molecule using segmental labeling is presented in Chapter 4. Having successfully produced a high molecular weight target for study, the next logical step is data acquisition. Hence, the final chapter on this theme, Chapter 5, describes TROSY methods for stru- ural studies.

Identification of desirable genotypes with traits of interest is discernible for making genetic improvement of crop plants. In this direction, screening of a large number of germplasm for desirable traits and transfer of identified traits into agronomic backgrounds through recombination breeding is the common breeding approach. Although visual screening is easier for qualitative traits, its use is not much effective for quantitative traits and also for those, which are difficult to score visually. Therefore, it is imperative to phenotype the germplasm accessions and breeding materials precisely using high throughput phenomics tools for challenging and complex traits under natural, controlled and harsh environmental conditions. Realizing the importance of phenotyping data towards identification and utilization of a germplasm as donors, global scientific community has exerted increased focus on advancing phenomics in crop plants leading to development of a number of techniques and methodologies for screening of agronomic, physiological, and biochemical traits. These technologies have now become much advanced and entered the era of digital science. This book provides exhaustive information on various aspects related to phenotyping of crop plants and offers a most comprehensive reference on the developments made in traditional and high throughput phenotyping of agricultural crops.

Proteomics is a multifaceted, interdisciplinary field which studies the complexity and dynamics of proteins in biological systems. It combines powerful separation and analytical technology with advanced informatics to understand the function of proteins in the cell and in the body. This book provides a clear conceptual description of each facet of proteomics, describes recent advances in technology and thinking in each area, and provides details of how these have been applied to a variety of biological problems. It is written by expert practitioners in the field, from industry, research institutions, and the clinic. It provides junior and experienced researchers with an invaluable proteomic reference, and gives fascinating glimpses of the future of this dynamic field. Written for: Libraries, institutes; scientists

ss-barrel outer membrane channel proteins (OMP) are useful as robust and flexible models or components in nanotechnology. Over the last decade biotechnological techniques allowed to expand the natural characteristics of OMPs by modifying their geometry and properties. The present book is oriented towards a broad group of readers including graduate students and advanced researchers. It gives a general introduction to the field of OMP based nano-component development as well as the state of the art of the involved research. On the example of the E. coli FhuA the transformation of an OMP into a tailored nano-channel will be outlined. An exhaustive description of the scientific strategy, including protein selection, analytical methods and "in-silico" tools to support the planning of protein modifications for a targeted application, consideration on the production of a custom made OMP, and an overview on technological applications including membrane/polymersome technology, will be provided.

The dynamic field of flavin and flavoprotein biochemistry has seen rapid advancement in recent years. This comprehensive two volume set provides an overview of all aspects of contemporary research in this important class of enzymes. Topics treated include flavoproteins involved in energy generation, signal transduction and electron transfer (including respiration); oxygen activation by flavoproteins; the biology and biochemistry of complex flavoproteins; flavin and flavoprotein photochemistry/photophysics as well as biotechnological applications of flavoproteins. Recent developments in this field include new structures (including those of large membrane-integral electron transfer complexes containing FMN or FAD), elucidation of the role of flavoproteins in cell signalling pathways (including both phototaxis and the circadian cycle) and important new insights into the reaction mechanisms of flavin-containing enzymes. This volume focusing on complex flavoproteins and physical methods is an essential reference for all researchers in biochemistry, chemistry, photochemistry and photophysics working on flavoenzymes.

The second edition covers a wide range of protein-protein interaction detection topics. Protein-Protein Interactions: Methods and Applications focuses on core technological platforms used to study protein-protein interactions and cutting-edge technologies that reflect recent scientific advances and the emerging focus on therapeutic discovery. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of necessary materials and reagents, step-by-step laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. These well-detailed protocols describe methods for identifying protein-protein interaction partners, analyzing of protein-protein interactions quantitatively and qualitatively, monitoring protein-protein interactions in live cells, and predicting and determining interaction interfaces. Authoritative and cutting-edge, Protein-Protein Interactions: Methods and Applications, Second Edition is a valuable resource that will enable readers to elucidate the mechanisms of protein-protein interactions, determine the role of these interactions in diverse biological processes, and target protein-protein interactions for therapeutic discovery.

Hands-on researchers describe in step-by-step detail 73 proven laboratory methods and bioinformatics tools essential for analysis of the proteome. These cutting-edge techniques address such important tasks as sample preparation, 2D-PAGE, gel staining, mass spectrometry, and post-translational modification. There are also readily reproducible methods for protein expression profiling, identifying protein-protein interactions, and protein chip technology, as well as a range of newly developed methodologies for determining the structure and function of a protein. The bioinformatics tools include those for analyzing 2D-GEL patterns, protein modeling, and protein identification. All laboratory-based protocols follow the successful Methods in Molecular Biology (TM) series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

A major direction in medical research leading to clinical applications targets the regulation of intracellular calcium and the various human diseases associated with an altered homeostasis of this global second messenger. These diseases include, for example: cardiomyopathy, inflammation, brain disorders, diabetes and cancer. In Calcium-Binding Proteins and RAGE: from Structural Basics to Clinical Applications,expert researchers in the field detail many of the methods which are now commonly used to study calcium binding proteins. These methods and techniques, such as calcium-measurements, screening methods, clinical chemistry, and therapy, are generally applicable to many other areas of basic and medical research as well as to diagnostics. 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,Calcium-Binding Proteins and RAGE: from Structural Basics to Clinical Applications underlines the diagnostic and clinical importance of this family of proteins in human diseases and as drug targets.