Electrochemical Analysis of Proteins and Cells presents the remarkable progress made over the years in the electrochemical analysis of proteins and cells, due to the rapid development of protein electrochemistry together with related technologies such as surface modification, molecular recognition, molecular assembly, and nanotechnology. As an interdisciplinary field combining electrochemistry, analytical chemistry, biochemistry, biophysics, biomedicine and material science, the electrochemical analysis of proteins and cells has attracted broad and extensive research interest. The main emphasis of this book is on the principles of electrochemical strategies and the practical utility of related detection systems, which is of great importance in all biological sciences, such as cell biology and molecular biology, as well as in biomedical fields like cancer research. This brief offers an up-to-date, easy-to-follow presentation of recent advances on the subject and can serve as a supplement for graduate-level courses in analytical chemistry, biochemistry, biophysics, biotechnology, biomedical engineering, etc. It may also help young scientists get an overview of this topic.

The book is devoted to expanding current views on the phenomena of protein functionality in food systems. Protein functionalities in foods have been the object ofextensive research over the last thirty to forty years and significant progress has been made in understanding the mechanism and factors influencing the functionality of proteins. The functionality of proteins is one of the fastest developing fields in the studies of protein utilization in foods. Currently, a broad spectrum of data related to protein functionality in food systems has been collected, however, much more needs to be known. In this volume, the most important functional properties offood proteins are presented: Protein solubility, water holding capacity and fat binding, emulsifying, foaming, and gelling properties as affected by protein source, environmental factors (pH, temperature, ionic strength) and protein concentration; Relationships between protein conformation, physicochemical properties, and functional properties; Protein functional properties as influenced by various food processing conditions, particularly heat treatment, dehydration, freezing and storage when frozen, extraction and other processes; Effects ofprotein modification on the enhancementofprotein functionality; Utilization ofvarious proteins in improving functional properties in food systems. Those aspects of protein functionality are presented which the author believes to be interesting and most important for protein utilization in food systems. The book is recommended to students and food scientists engaged in food protein research and food industry research, and development scientists. Table ofContents Introduction 1 References 5 Chapter 1 Solubility ofProteins. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 1. 1 Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 1. 1. 1 Factors Affecting Solubility ofProteins. . . . . . . . . . . . . . . . . . . . . . . .

Research in the pharmaceutical industry today is in many respects quite different from what it used to be only fifteen years ago. There have been dramatic changes in approaches for identifying new chemical entities with a desired biological activity. While chemical modification of existing leads was the most important approach in the 1970s and 1980s, high-throughput screening and structure-based design are now major players among a multitude of methods used in drug discov ery. Quite often, companies favor one of these relatively new approaches over the other, e.g., screening over rational design, or vice versa, but we believe that an intelligent and concerted use of several or all methods currently available to drug discovery will be more successful in the medium term. What has changed most significantly in the past few years is the time available for identifying new chemical entities. Because of the high costs of drug discovery projects, pressure for maximum success in the shortest possible time is higher than ever. In addition, the multidisciplinary character of the field is much more pronounced today than it used to be. As a consequence, researchers and project managers in the pharmaceutical industry should have a solid knowledge of the more important methods available to drug discovery, because it is the rapidly and intelligently combined use of these which will determine the success or failure of preclinical projects.

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.

-Lignin Structure, Properties, and Applications By H. Hatakeyama, T. Hatakeyama -Tensile Mechanics of -Helical Coil Springs By A. Ikai -Bioactive Polymer/Hydroxyapatite (Nano)composites for Bone Tissue Regeneration By K. Pielichowska, S. Blazewicz"

The highly structured eucaryotic cell with its complex division of biochemical labour requires a distinct protein complement in each cellular structure and compartment. Nuclear coded and cytosolically synthesized polypeptides are specifically sorted to every corner of the cell in a post- or co-translational manner. The presence of separate genomes and protein translation machineries in plastids and mitochondria requires further coordination not only on the transcriptional, translational but also most likely on the protein import level. Numerous different protein transport systems have developed and coexist within plant cells to ensure the specific and selective composition of every sub-cellular compartment. This volume summarizes the current knowledge on protein trafficking in plant cells. Aside from the fundamental aspects in cell biology of how specific pre-protein sorting and translocation across biological membranes is achieved, a major focus is on transport, modification and deposition of plant storage proteins. The increasing use of plants as bioreactors to provide custom-designed proteins of different usage requires detailed understanding of these events. This text is directed not only at students and professionals in plant cell and molecular biology but also at those involved in horticulture and plant breeding. It is intended to serve as a text and guide for graduate-level courses on plant cell biology and as a valuable supplement to courses in plant physiology and development. Scientists in other disciplines who wish to learn more about protein translocation in plants will also find this text an up-to-date source of information and reference.

The book addresses the most recent developments in structural and functional proteomics underlying the recent contributions given in these areas by our laboratory to the instrumentations, the methods and the procedures as mutuated from the nanoscale sciences and technologies. These developments introduced in the last few years make now possible protein massive identification (mass spectrometry and biomolecular arrays down to nanoamounts) and protein structural characterization in solution and in crystals down to the atomic scale to an extent and to a degree so far unmatched. Emphasis is placed in the growth by nanobiofilm template of protein crystals of any type and size from millimeter to micron, leading in combination with microfocus synchrotron technology and atomic force microscopy to the definition of a new field called nanocrystallography. The few useful examples being shown, concerning yet structurally unsolved proteins, point this very promising approach nanotechnology-based in structural proteomics using highly focused X-rays. This has not to be confused with the important study of nanocrystals, both organic and inorganic, and novel diamond like nanocomposite materials and devices having 3D protein crystals as matrices to be equilibrated with nanoparticles/gold/silver to be utilized in the most diversified electronic applications here also summarized. vii Acknowledgments We are particularly grateful to Giuseppe Zanotti at the University of Padova for his fundamental collaboration during all the crystallographic studies.

PAS proteins control numerous physiological and developmental events, and span phylogeny from bacteria to man. Bacterial and plant PAS proteins act as sensors of environmental stimuli, including light, oxygen, and energy status. Not surprising, given these roles, there is intense investigation of the roles of bHLH-PAS proteins in issues of human health including: (1) cancer induction, (2) cancer growth and vascularity, (3) birth defects, including Down syndrome, (4) appetite control and obesity, (5) sleep rhythm disorders, and (6) mental health disorders such as social interactions and learning. PAS proteins encompass many fields of biology, and scientists who work in these fields (circadian rhythms, oxygen regulation, toxin metabolism, bacterial sensors, and development) are an audience, particularly those who actively work on PAS proteins and researchers interested in transcriptional control, signal transduction, and evolution.

Physics and the life sciences have established new connections within the past few decades, resulting in biological physics as an established subfield with strong groups working in many physics departments. These interactions between physics and biology form a two-way street with physics providing new tools and concepts for understanding life, while biological systems can yield new insights into the physics of complex systems. To address the challenges of this interdisciplinary area, The Physics of Proteins: An Introduction to Biological Physics and Molecular Biophysics is divided into three interconnected sections. In Parts I and II, early chapters introduce the terminology and describe the main biological systems that physicists will encounter. Similarities between biomolecules, glasses, and solids are stressed with an emphasis on the fundamental concepts of living systems. The central section (Parts III and IV) delves into the dynamics of complex systems. A main theme is the realization that biological systems, in particular proteins, do not exist in unique conformations but can assume a very large number of slightly different structures. This complexity is captured in the concept of a free energy landscape and leads to the conclusion that fluctuations are crucial for the functioning of biological systems. The final chapter of this section challenges the reader to apply these concepts to a problem that appears in the current literature. An extensive series of appendices (Part V) provide descriptions of the key physical tools and analytical methods that have proven powerful in the study of the physics of proteins. The appendices are designed to be consulted throughout the section on protein dynamics without breaking the deductive flow of the logic in the central section of the book.

From the basic science to potential and approved clinical applications the most recent data in the rapidly growing field of bone morphogenetic proteins (BMPs) are summarized in this topical volume. Distinguished scientists present reviews on a range of scientific topics, including biochemistry, biology, molecular biology and preclinical animal studies on spinal fusion, cartilage repair, craniofacial and dental reconstruction using BMPs, as well as approved clinical applications in human bone non-unions.

This book provides a resource not only for experts in the field, but also for undergraduate students, newcomers and clinicians worldwide, given that the use of BMPs in orthopedic reconstruction has been already approved in Europe, Australia, Canada and the USA.

This thesis describes research into the mode of function, inhibition, and evolution of the ribosomal catalytic center, the Peptidyl Transferase Center (PTC)--research that has already led to attempts at improving PTC antibiotics. The PhD candidate carried out two parallel studies. One using a combination of X-ray crystallography, biochemistry, molecular biology, and theoretical studies to obtain crystal structures of ribosomal particles with antibiotics that target the PTC, revealing the modes of action, resistance, cross-resistance and discrimination between ribosomes of eubacterial pathogens and eukaryotic hosts. In the second parallel study, the candidate synthesized a ribosomal substructure--one that may represent the minimal entity capable of catalyzing peptide bond formation--shedding light on the origin of the ribosome itself.

The present book relates to the scientific records of a workshop held in Patras, Greece, in June 1989, under the auspices and with financial support of the European Economic Communities (Concerted Action EUROBIOMAT - Hemocompatibility - of the Medical Research Programme, Project: 11.1.212). This concerted action promotes the collaboration on science and technology on the particular field of hemocompatible biomaterials: exchange of experts, scholarships and scientific workshops within the EC-member countries and COST countries such as Sweden, Finland, Turkey, Switzerland. The first part of this monography refers to the oral presentations of the par ticipants. The second part gives the book its unique character: the scientific discussion on updated aspects of protein adsorption of synthetic polymers in contact with blood. This second part is subdivided into nine chapters where specific topics were discussed freely, open-minded and even controversially. This book intends to elucidate recurrent questions concerning the initial event when blood contacts artificial surfaces. Young investigators will consider this book to be appropriate to get familiar with the scientific background and the most relevant techniques and methods."

Using genome sequencing, one can predict possible interactions among proteins. There are very few titles that focus on protein-protein interaction predictions in bacteria. The authors will describe these methods and further highlight its use to predict various biological pathways and complexity of the cellular response to various environmental conditions. Topics include analysis of complex genome-scale protein-protein interaction networks, effects of reference genome selection on prediction accuracy, and genome sequence templates to predict protein function.

The standard protocols for the purification of all known cytoskeleton proteins are presented in this manual. Proteins are listed alphabetically and each protocol follows a common format. Thus, the manual provides a quick and easy reference to all relevant procedures for cytoskeleton protein purification.
The isolation procedure for each protein is shown in a clear flowchart, while the source of the protein, equipment and material needed, a list of suppliers, standard references, accession No. of sequences as well as further relevant facts and practical tips are given on a separate page.

In the ten years since the publication of the first edition, great advances in fluorescent labeling, optics, and sample preparation have significantly improved the imaging capability of microscopy, allowing for a continual refinement of our understanding of the cytoskeleton as a dynamic synergy of components. In Cytoskeleton Methods and Protocols, Second Edition, internationally renowned experts present techniques which reflect many of the recent technological advances in experimental tools for cytoskeleton research with emphasis on animal, plant, protist, and fungal model systems. This cutting-edge volume contains methods for live-cell imaging, fluorescence microscopy, electron microscopy, analysis of cell and organelle motility, isolation of cytoskeleton components, and proteomics, amongst other topics. As a volume in the highly successful Methods in Molecular Biology series, chapters incorporate introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes that provide unpublished technical information on troubleshooting and avoiding known pitfalls.

Up-to-date and comprehensive, Cytoskeleton Methods and Protocols, Second Edition serves as an ideal guide to scientists who wish to continue this fruitful and important biological research."

A NATO Advanced Study Institute on "Signal Transduction and Protein Phosphorylation" was held to overview recent developments in this area. The participants in the Institute dealt with protein phosphorylation as the most prevalent mode of regulation of cellular processes. First, methods needed to analyze the complex cascade systems involved were reviewed, including protein sequencing, crystallo- graphy, characterization and isolation of membrane proteins, use of monoclonal or polyclonal antibodies and application of fluorescent probes. In great detail the x ray crystallographic structure of glycogen phosphorylase was presented. This enzyme is located at the end of a signal cascade triggered by the hormonal activation of the membrane-bound adenylate cyclase. The interaction of the hormone/receptor with the catalytic subunit of the adenylate cyclase involves GTP-binding proteins. The function of these recently detected intermembrane coupling factors were reviewed, as weIl as the structure and properties of various protein kinases. 2 Major emphasis was placed on Ca + as a second messenger, its metabolism, mechanism of release and uptake from intracellular stores and its role on cell motility and muscle contraction. Two classes of protein phosphatases were discussed. They differ in their subunit structure and substrate specificity and are subject of a highly complex regulatory mechanism as yet not fully under- stood.

With the invitation to edit this volume, I wanted to take the opportunity to assemble reviews on different aspects of circadian clocks and rhythms. Although most c- tributions in this volume focus on mammalian circadian clocks, the historical int- duction and comparative clocks section illustrate the importance of various other organisms in deciphering the mechanisms and principles of circadian biology. Circadian rhythms have been studied for centuries, but only recently, a mole- lar understanding of this process has emerged. This has taken research on circadian clocks from mystic phenomenology to a mechanistic level; chains of molecular events can describe phenomena with remarkable accuracy. Nevertheless, current models of the functioning of circadian clocks are still rudimentary. This is not due to the faultiness of discovered mechanisms, but due to the lack of undiscovered processes involved in contributing to circadian rhythmicity. We know for example, that the general circadian mechanism is not regulated equally in all tissues of m- mals. Hence, a lot still needs to be discovered to get a full understanding of cir- dian rhythms at the systems level. In this respect, technology has advanced at high speed in the last years and provided us with data illustrating the sheer complexity of regulation of physiological processes in organisms. To handle this information, computer aided integration of the results is of utmost importance in order to d- cover novel concepts that ultimately need to be tested experimentally.

The Discovery of Ribonuclease P and Enzymatic Activity of Its RNA Subunit Sydney Brenner and Francis H. C. Crick had a specific project in mind when they offered Sidney Altman a position in their group in 1969 to conduct postdoctoral research at the Medical Research Council Laboratory of Molecular Biology (LMB) in Cambridge, England. At the time, an intense international competition was on- ing in as many as a dozen labs to determine the three-dimensional structure of tRNA. At the LMB, Aaron Klug was attacking the structure by crystallographic analysis with Brian F. C. Clark providing large amounts of purified phenylalanine tRNA. (Eventually, Aaron announced his empirically determined 3-D structure of yeast phenylalanine tRNA, a structure that is generally common to tRNAs, due in part to several conserved, novel three-way nucleotide interactions. ) Concurrently, Michael Levitt, a Ph. D. student of Francis, was visually scrutinizing the cloverleaf secondary structure of the 14 tRNA sequences known at the time. Levitt was searching for nucleotide covariation in different parts of the molecules that were conserved in the 14 sequences known at the time. He identified a possible covariation of an apparent Watson-Crick pairing type between the residues at position 15 from the 5' end of the tRNA and residue 48. This association implied these parts of the tRNA, namely the D loop containing residue 15 and the 5' end of the T stem-adjoining residue 48, folded on one another in a tertiary structure shared by different tRNAs.

The feld of proteomics moves rapidly. New methods, techniques, applications, standards, models and software appear almost on a daily basis. Accompanying this are plenty of texts on the experimental side of the feld and a few appearing on the informatic and data analysis side. This latterly includes one in the Methods in Molecular Biology series tackling the specifc analysis of "Mass spectrometry data in proteomics" in MMB vol. 376. This current collection builds on this, but takes a broader view of proteome data analysis covering data analysis essentials, but also the databases and data models, as well as practical consid- ations for analysing database search results, annotating genomes, and speeding up searches. It also digs deeper into some topics, such as decoy database searching and aspects of signal processing in proteomic mass spectrometry. The aim of the volume is to provide the reader with a mix of reviews and methodology chapters, which build from the essentials of database searching in proteomics, on through specifc data processing challenges to databases, data standards and data models.

Hunger is the world problem Nr 1, overshadowed by an uncontrollable explosion of the human population all over our planet. Lack of food has been one of the most primitive dangers, which animal life had to face at every stage of its evolution. The living body developed in the course of this evolution special emergency reactions against this danger, which is characterised by a lack of food calories, a lack of nitrogen in the form of proteins, a lack of vitamins and oligo elements. Based on an intricate physiological defense pattern man can support complete starvation up to one month, by using up the substance of less important organs in order to maintain the functional matrix of the important organs, mainly the brain and the nervous system. This regulation and the pattern of its mechanisms are of great interest to the physiologist who is aware that they are also responsible for the maintenance of life among millions of human beings who desperately live in a state of permanent hunger. The most serious problem in many developing countries is not the supply of calories (mainly carbohydrate calories) or the supply of vitamins and oligo elements, but the supply of a sufficient amount of protein in order to overcome the protein-calorie malnutrition. This problem must be considered as the most urgent one among all the other problems in the fight against hunger.

A number of apparently unrelated phenomena in biological systems (e.g., biopolymer aggregation, cell-cell interactions, ion transport across membranes) arise from the special properties of charged surfaces. A sym posium entitled "Electrical Double Layers in Biology," which took place at the Toronto meeting of the Electrochemical Society, 12-17 May 1985, focused on the common features of these phenomena. The papers presented at that symposium are collected here and they illustrate ways in which an under standing of electrical double layers can elucidate a problem in Biology. An example of this approach can be seen from the paper I presented on ion transport and excitation, where the "unusual" ion flows during nerve excitation are actually expected if one includes the effects of electrical double layers at membrane surfaces. Furthermore, the selectivity of the ion channels in these membranes can be better understood on this basis. Other presentations account for such observations as the changes in spacing between muscle proteins during contraction, the interactions of red cells to form rouleaux, the electrical properties of algal cell membranes, electrokinetic potentials during blood flow in arteries, etc. I trust that these papers will indicate the value of electrochemistry in the study of biological systems, an area of research usually called Bioelectrochemistry, and will encourage biologists to use these ideas when approaching related problems."

A solution to the protein folding problem has eluded researchers for more than 30 years. The stakes are high. Such a solution will make 40,000 more tertiary structures available for immediate study by translating the DNA sequence information in the sequence databases into three-dimensional protein structures. This translation will be indispensable for the analy sis of results from the Human Genome Project, de novo protein design, and many other areas of biotechnological research. Finally, an in-depth study of the rules of protein folding should provide vital clues to the protein fold ing process. The search for these rules is therefore an important objective for theoretical molecular biology. Both experimental and theoretical ap proaches have been used in the search for a solution, with many promising results but no general solution. In recent years, there has been an exponen tial increase in the power of computers. This has triggered an incredible outburst of theoretical approaches to solving the protein folding problem ranging from molecular dynamics-based studies of proteins in solution to the actual prediction of protein structures from first principles. This volume attempts to present a concise overview of these advances. Adrian Roitberg and Ron Elber describe the locally enhanced sam pling/simulated annealing conformational search algorithm (Chapter 1), which is potentially useful for the rapid conformational search of larger molecular systems."

The Ottawa '88 meeting of the International Society for Oxygen Transport to Tissue attracted a record number of participants and presentations. We were able to avoid simultaneous sessions and still keep the scientific program to four days by using poster sessions followed by plenary debate on each poster. To paraphrase the British physicist David Bohm, we tried to avoid an ordinary discussion, in which people usually stick to a relatively fixed position and try to convince others to change. This situation does not give rise to anything creative. So, we attempted instead to establish a true dialogue in which a person may prefer and support a certain point of view, but does not hold it nonnegotiab1y. He or she is ready to listen to others with sufficient sympathy, and is also ready to change his or her own view if there is a good reason to do so. Our Society is in its "teen" years, and there are even some arguments about its exact age. Many newer members have raised questions concerning the history of the Society. For this reason, I have asked one of the "founding fathers," D. Bruley, to prepare a brief account of the birth and early history of the Society which appears on the following page.

Trying to address the entire field of presynaptic modulation of neurotransmitter release is a rather daunting undertaking, one that is well beyond the scope of this book. In addition, studies of release modulation, particularly from a biochemical standpoint, have been the subjects of several extensive reviews, meetings, and books (Langer, 1978; Chesselet, 1984; Wessler, 1989; Kalsner and Westfall, 1990), which provide an essential introduction to this subject. What we have focused on, however, are several specific aspects of release modulation that perhaps have not been as extensively discussed. First, we felt that it was important to focus on modulation in the central nervous system; much of the work that has been done in the past has emphasized the peripheral nervous system (e. g. , the autonomic nervous system and the neuromuscular junction), in part because such preparations are more amenable to study. However, it is becoming clear that modulation of release is, if anything, more important in the central nervous system than in the periphery, and that virtually every transmitter system that has been studied shows some type of release modulation. The other way in which we have restricted the scope of this volume has been to try to emphasize studies in which functional (primarily electrophysiological) measures of transmitter release have been used rather than direct biochemical measures of release, and to explore the ways in which release modulation affects the normal physiological function at synapses.