Application of NMR and Molecular Docking in Structure-Based Drug Discovery, by Jaime L. Stark and Robert Powers NMR as a Unique Tool in Assessment and Complex Determination of Weak Protein-Protein Interactions, by Olga Vinogradova and Jun Qin The Use of Residual Dipolar Coupling in Studying Proteins by NMR, by Kang Chen und Nico Tjandra NMR Studies of Metalloproteins, by Hongyan Li and Hongzhe Sun Recent Developments in 15N NMR Relaxation Studies that Probe Protein Backbone Dynamics, by Rieko Ishima Contemporary Methods in Structure Determination of Membrane Proteins by Solution NMR, by Tabussom Qureshi and Natalie K. Goto Protein Structure Determination by Solid-State NMR, by Xin Zhao Dynamic Nuclear Polarization: New Methodology and Applications, by Kong Hung Sze, Qinglin Wu, Ho Sum Tse and Guang Zhu

-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 Proceedings of the Fourth International Metallothionein Meeting (MT-97) feature the latest research on metallothionein. The book covers a broad range of topics which provide important information for both basic and clinical investigators. The selected 94 articles in this book are written by the leading scientists in the field around the world. This is an increasingly important, multi-disciplinary area of study that has benefitted from recent advances in concepts and methodologies from other fields.

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.

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.

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."

LIST OF PARTICIPANTS 11 THERMOPHILIC HYDROLYTIC ENZYMES (PROTEINASES, AMYLASES) 17 Thermal Stability of Homologous Neutral Metalloendopeptidases in Thermo philic and Mesophilic Bacteria: Structural Considerations M. K. PANGBURN, P. L. LEVY, K. A. WALSH and H. NEURATH 19 The Structure and Stability of Thermolysin L. H. WEAVER, W. R. KESTER, K. F. TEN EYCK and B. W. MATTHEWS 31 Studies of the Inhibition of Thermolysin J. FEDER, N. AUFDERHEIDE and B. S. WILDI 41 Effect of EDTA on the Conformational Stability of Thermolysin A. FONTANA, E. BOCCU and F. M. VERONESE 55 Role of a Sulfhydryl Group in the Structure and Function of Alkaline Proteases from a Thermophilic Actinomycete K. MIZUSAWA and F. YOSHIDA 61 Partial Characterization of a Thermophilic Actinomycete Rennin S. LAXER, A. PINSKY and B. BARTOOV 67 Amylase Activity and Stability at High and Low Temperature Depending on Calcium and other Divalent Cations W. HEINEN and A. M. LAUWERS 77 Role of Calcium Ion in the Thermostability of a-Amylase Produced from Bacillus stearothermophilus K. YUTAN I 91 THERMOPHILIC DEHYDROGENASES 105 Thermophilic Glyceraldehyde-3-P Dehydrogenase R. E. AMELUNXEN and R. SINGLETON JR. 107 Glyceraldehyde 3-Phosphate Dehydrogenase from an Extreme Thermophile, Thermus aquaticus 121 J. D. HOCKING and J. I. HARRIS Thermal Properties of Glyceraldehyde 3-Phosphate Dehydrogenase from Escherichia coli A. FONTANA, C. GRANDI, E. BOCCU and F. M. VERONESE 135 Comparative Conformational Properties of Thermophilic and Mesophilic 6-Phosphogluconate Dehydrogenase F. M. VERONESE, C. GRANDI, E. BOCCU and A."

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.

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."

Within the past two decades, extraordinary new functions for the nucleolus have begun to appear, giving the field a new vitality and generating renewed excitement and interest. These new discoveries include both newly-discovered functions and aspects of its conventional role. The Nucleolus is divided into three parts: nucleolar structure and organization, the role of the nucleolus in ribosome biogenesis, and novel functions of the nucleolus.

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.

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 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."

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 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.

The book is dedicated to the topical area of biology and medicine and the role of stress proteins HSP70 in the regulation of intracellular protein homeostasis, signaling transduction and cell protection. The book is divided into chapters, which describe the discovery of HSP70 and its molecular structure, the mechanism of the synthesis and function in normal and damaged cells, examine the role of HSP70 in immunity, cancerogenesis, aging, Alzheimer's disease and cardiac surgery. In this book, the author looks at HSP70 as a factor which prevents the transformation of homeostasis mechanisms of intracellular proteins into a link in the pathogenesis of a disease.

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.

Recent publications have addressed specific aspects of the endothelins, such as their roles in disease or the importance of endothelin receptors. However, in this book the entire field of endothelins is covered. This includes the pathways of endothelin production and their regulation, the local and systemic actions of endothelins, receptors for the endothelins and the signalling pathways employed, and the involvement of endothelins in a range of diseases. Attention is also paid to the development through chemistry, pharmacology and toxicology of endothelin antagonists and endothelin-converting enzyme inhibitors, with mention of all the important members of these drug classes. This leads to well-rounded discussions of the potential therapeutic benefit of endothelin inhibitors.

A century has already passed since FRIEDRICH MIESCHER, working at Strasbourg and Basel, began his study of protamine, one of the basic nuclear proteins of cells. It was first established by KOSSEL that protamine represents the simplest known protein. In the conviction that research into the nature of protamine would shed light on that of other typical proteins, a group of researchers in Germany followed MIESCHER and laid the foundations of protein chemistry. A general view of prot amines was thus built up by KOSSEL, working at Strasbourg, Berlin, Marburg an der Lahn, and Heidelberg, FELIX at Heidelberg, Munich, and Frankfurt am Main, and WALDSCHMIDT-LEITZ at Prague and Munich. Concepts and techniques established by these studies have been widely utilized for research on other typical proteins. The revolutionary advances in chemical and physical techniques after W orId War II extended the sphere of research to Tokyo in the Far East. Prof. FELIX' visit in 1955 greatly encouraged our research group in Tokyo. His death in August 1960 constituted a sad loss to protein chemistry and stimulated our group to assume responsibility for carrying on the studies. In the following decade we in Tokyo have been able to add a new development to the results on the chemical structure of protamines accumulated by the Eurqpean researchers over a period of about fifty years."

This volume contains the proceedings of the International Conference on Prostaglandins and Lipid Metabolism in Radiation Injury held in Rockville, Maryland, on October 2-3, 1986. Over 200 persons from eight countries attended the program, which consisted of 24 oral presentations and 38 poster presentations. Forty-two of those presentations have been included in this volume. The conference was sponsored by the Armed Forces Radiobiology Research Institute, located in Bethesda, Maryland. The effects of radiation on lipid synthesis and membrane damage are aptly summarized in the first five chapters. These chapters describe the effects of radiation on lipid peroxidation of model membranes, and the role of lipid composition in mammalian cell death and in bacterial radio- and thermosensitivity. In bacteria, lipid peroxidation is not essential for radiation-induced cell death. One of the key points of the conference was the paradoxical nature of the radiobiology of eicosanoids. On the one hand, eicosanoids are mediators of damage; on the other, they are radioprotective agents. It is clear from the literature and from the data presented at the conference that both of these properties may also be observed as a consequence of radiotherapy. Some studies indicate that nonsteroidal anti-inflammatory drugs may minimize or prevent certain radiation induced damage, but other studies show no positive effect."

The pleasant community of Limone suI Garda provided outstanding hospitality for a second NATO ARW dealing with apolipoprotein variants, which are natures clues for the discovery of the physiological roles of apolipoproteins in lipoprotein metabolism in normal subjects and patients with specific dyslipoproteinemias. Limone, the site of discovery of the first human apolipoprotein mutant, apoA-I-Milano, provided a brilliant sunny spring venue for more than 50 participants from both sides of the ocean. The attendance at the colorful opening ceremony of the ARW was one of the largest on record. Two members of the Italian government, the Secretaries of Health and the Navy, gave the welcoming addresses. Six television networks, two with national audiences, covered the international workshop. The Limone oracles provided a montage of insights gleamed from the eyes of the clinican, the biochemist, and the molecular biologist. The cumulative information on the molecular defects in lipoprotein metabolism reviewed by this diverse group of investigators provided an ever expanding horizon of new knowledge in this fast moving and some times perplexing field. Clinical vignettes were presented on patients from throughout the world including Canada (Connelly), Turkey (Schmitz), and France (Infante) detailing the clinical sequelae of a defect in a specific apolipoprotein. The clinical importance of Lp(a), a lipoprotein relegated almost to obscurity for many years, has now taken v center stage.

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.