Subcellular Biochemistry – serie

This volume and future ones in the series, which is now under new editorship. depart from the pattern established by previous volumes in containing chapters loosely related to one another. It is thus appropriate that each volume should have a relevant subtitle. In the present volume the underlying theme is immu­ nology and its application to the study of a number of different biochemical systems. Almost inevitably, the study of the cell-surface antigens and glycoproteins of a number of different tissues tends to dominate the contents, but considerable emphasis is also placed on subcellular organelles and enzymes. Undoubtedly, it is the impact of readily available monoclonal antibodies that dominates work in the field. The epitope specificity provided by monoclonal antibodies gives the biochemist a powerful lever with which to probe the location, structure, and function of cellular protein and glycoprotein antigens. A number of the chapters included in this volume have a strong biomedical emphasis. This is indeed intentional, as it is now the policy of the series to cover the more applied aspects ofthe subject, while in no way neglecting more fundamentaltopics in subcellular biochemistry.

Cellular virology has made tremendous advances in the past decade due to the availability and application of new immunological techniques together with the vast range of biochemical techniques and the continued impact of transmission electron microscopy. The chapters contained in this volume provide significant coverage of the subject of cellular virology as a whole. Considerable overall emphasis is placed upon the membrane biochemistry of viral proteins and glycoproteins within the infected cell. In the opening chapter Edouard Kurstak and his colleagues provide a useful survey on the detection of viral antigens and antibodies by immunoassays. This chapter, with its emphasis on the important role of immunology in present-day virology, sets the scene for the volume. Following this is an exciting presentation from Stefan Hoglund and his colleagues on ISCOMs and immunostimulation with viral antigens. This unique approach is already proving to be of value, particularly in animal virology. A somewhat pharmacological diversion appears in Chapter 3, by Kazukiyo Onodera and his colleagues, in which the biological activity of the damavaricin C derivatives is discussed. This chapter provides a link between the biochemical and the chemotherapeutic approach in cellular virology. Yet another specialist area is covered in Chapter 4 by Otto Schmidt and hnke Schuchmann-Feddersen, who discuss the role of virus-like particles in para­ site-host interactions of insects. Contributing a strong biomedical emphasis to the volume is the provocative chapter by Abraham Karpas on human leukemia and retroviruses.

Corpora non agunt nisi fixata. This old saying of Ehrlich's describing the physi- ological role of receptors and their ligands might be paraphrased into Corpora non ambulant nisi fixata when considering lipid transport between and within cells. Volume 16 of Subcellular Biochemistry is intended to bring the reader up to date with this young field. Indeed, lipid transfer proteins have only recently become the subject of a more systematic study. In this book the current status and the emerging trends are discussed. Chapters cover protein-mediated transfer of fatty acids, phospholipids, phosphatidylinositol, glycolipids, dolichol, retinoids, and cholesterol in animal, plant, yeast, and other eukaryotic cells. Details are included of the study of lipid transport proteins by means of fluorescent phos- pholipid analogues and of the lipid transfer proteins as probes of membrane structure and function, as well as spontaneous lipid transfer as it occurs between biological membranes. Some of the chapters should be read in conjunction with Volume 13 of this series, devoted to fluorescence studies on biological mem- branes, in particular Chapter 2 (Somerharju et al.) concentrating on studies in which fluorescent phospholipid analogues have been used. Chapter 10 (Bill- heimer and Reinhart), dealing with cholesterol trafficking, should be compared with Chapter 12 of Volume 13 (Van Blitterswijk), pointing to the existence of a preferential association of cholesterol with sphingomyelin, which drags choles- terol to the plasma membrane. In one chapter (Chapter 8: Van Dessel et al.

Volume 18 of the Subcellular Biochemistry series, which specializes in various aspects of the biochemistry of the intracellular parasites, was initially proposed by Jose Luis Avila and strongly supported by myself, as Series Editor of Sub­ cellular Biochemistry. Considerable assistance was received from Professor Frank Wunderlich (University of Dusseldorf) and more particularly from Dr. Michael Miles (London School of Hygiene and Tropical Medicine) during the compilation of the list of possible chapters. Our initial aim was to present a comprehensive survey of this broad field of study. Although some interesting topics have been lost due to authors backing out late in the production schedule of the book, the manuscripts that were submitted have provided a useful over­ view of the subject, with notable strength within the field of Leishmania. The 13 chapters of the book have been grouped according to subject. The first five chapters deal with Leishmania and are followed by two chapters on Try­ ponosoma cruzi, two on the malarial parasites, and two on the Coccidia. The fmal two chapters cover the Microsporidia and chemotherapy, respectively.

The present volume is the first attempt to bring together, in a comprehensive and comparative manner, the vast knowledge accumulated on the mycoplasma cell membranes. The state-of-the-art coverage includes physical studies of lipid organization and dynamics in mycoplasma membranes, the role of cholesterol in mycoplasma membranes, spiralins, and eight other subjects.

In step with the surge of interest in the endoplasmic reticulum, the current volume takes an integrated look at this functionally diverse organelle. Coverage includes protein translocation and export, lipid metabolism, antigen presentation, and many other subjects, gleaned from such diverse fields as cell biology, enzymology and membrane biochemistry, immunology, and signal transduction.

This latest volume summarizes the current state of knowledge of membrane assembly. It covers subjects germane to most membranes as well as to membranes of different organelles for prokaryote, plant, and animal cells.

Here, researchers review the latest breakthroughs in protein research. Their contributions explore emerging principles and techniques and survey important classes of proteins that will play key roles in the field's future. Articles examine the possibility of a Boltzman-like distribution in protein substructures, the new technique of Raman spectroscopy, and compact intermediate states of protein folding. This well-illustrated volume also features coverage of proteins that bind nucleic acids.

Experts investigate the biochemical and biomedical aspects of cholesterol, addressing its metabolism in normal and disease states. They discuss a broad range of topics, including key steps in the cholesterol biosynthetic pathway, and the role of cholesterol in cancer, atherosclerosis, and diseases of the nervous system. The book's comprehensive coverage also includes the pathological consequences and potential therapies for various disease states, and the development of new anti-atherogenic drugs.

This volume provides a detailed review of research covering different modes of interaction between microbes and plants. Included among the interactions examined are: the symbiotic association of rhizobium, azorhizobium and frankia with different plant systems; disease resistance strategies for plants; the role of the agrobacterium system in the formation of tumours, and oligosaccharides produced as a result of the degradation of plant microbial extracellular matrix.

The subject for a volume on the fat-soluble vitamins needs no justification considering the importance of this group of nutrients and the rate of expan­ sion of our knowledge of its role in cell biology, genetics, and disease. The level of our understanding has clearly moved from knowing what fat­ soluble vitamins do to how they perform their functions. Hand in hand with a knowledge of their molecular mechanisms of action is the recognition that vitamins are used sparingly, and regeneration processes operate in certain cases to recycle vitamins from their metabolites. We have divided the volume into alphabetical sections beginning with vitamin A and the carotenoids through vitamins D, E, F, and K, and ending with coenzyme Q. The contributors are all acknowledged experts in their particular fields and have made significant contributions to published research results. All have worked assiduously to deliver the product of their labors on a restricted time scale and to provide the most up-to­ date information on their respective topics. We are truly grateful for their indulgence.

This text looks at subcellular chemistry, particularly at intermediate filaments. Topics include fish intermediate filament proteins in structure, evolution, and function, and lessons from keratin transgenic and knockout mice.

It has been 15 years since the first report on the isolation of anti-Gal from human serum and the demonstration that this antibody is the most prevalent antibody in humans (Galili et al. , ]. Exp. Med. 160: 1519, 1984). Subsequent interdisciplinary studies in immunology, carbohydrate biochemistry, molecular biology, and evo- lution demonstrated the highly restricted specificity of anti-Gal for the carbohy- drate epitope Gal al-3Galpl-4GIcNAc-R, (termed here the a-gal epitope), the unprecedented evolutionary pattern of distribution of a-gal and anti-Gal in mam- mals, and explained the evolution of this antigen and antibody by analysis of the a 1 ,3galactosyltransferase gene, the gene that encodes the enzyme that synthesizes the a-gal epitope. These studies have suggested that a major selection process that occurred in the course of evolution of ancestral Old World primates resulted in the inactivation of the a1 ,3galactosyltransferase gene and the subsequent appearance of anti-Gal in these primates. Other studies in immunoparasitology have demon- strated the possible physiologic significance of anti-Gal in protection against cer- tain parasitic infections.Major scientific attention was focused on a-gal and anti-Gal with the real- ization in the early nineties that the interaction between this antigen and antibody is the major obstacle to xenotransplantation. The success of immunosuppressive drugs, in the last two decades of the 20th century, in preventing allograft rejection, has raised hopes for cure in many patients in need of organ transplant. Because of limited supply of allografts, only 20% of patients receive the needed organ.

Strategies of Bacterial Interaction with Eukaryotic Cells *Tobias A. Oelschlaeger and Jorg Hacker 1. BENEFICIAL BACTERIAL-HOST INTERACTIONS Already during birth and soon thereafter mammals are colonized by bacte- ria belonging to the resident microbial flora. Cutaneous and mucosal sur- faces and the gastrointestinal tract are the areas which become colonized. These indigenous or autochthonous bacteria have a variety of beneficial effects on their hosts. They play a protective role by bacterial antagonism in fighting infections (Hoszowski and Truszczynski, 1997; Hentges, 1979). Pro- duction of vitamin K is another essential contribution of the resident microbial flora to the health of the host (Hill, 1997). Even more important, studies with germ-free animals demonstrated the involvement of the microbial flora on the development of the immune system. Such animals have underdeveloped and relatively undifferentiated lymphoid tissues and low concentrations of serum immune globulins ( Cebra et at., 1998). They TOBIAS A. OELSCHLAEGER and JORG HACKER Institut filr Molekulare lnfektionsbiologie, Universitiit Wiirzburg, 97070 Wiirzburg, Germany.*Corresponding author; Phone: (0)931-312150; FAX: (0)931-312578; E-mail: t.oelschlaeger@mail.uni-wuerzburg.de xxix Tobias A. Oelschlaeger and Jorg Hacker also show defects in specific immune responsiveness and in nonspecific resistance induced by endotoxin, which may account for their lowered resis- tance. A more typical example of symbiotic interaction of bacteria with a host are bacteria like Ruminococcus in the gut of ruminants, essential for degradation of cellulose (Hobson, 1988). The closest benefical bacterial-host interactions are those of intracellular symbiotic bacteria and their host cells.

Membrane fusion and targeting processes are tightly regulated and coordinated. Dozens of proteins, originating from both the cytoplasm and membranes are involved. The discovery of homologous proteins from yeast to neurons validates a unified view. Although much is known about the interfering proteins, the events occurring when two lipid bilayers actually fuse are less clear. It should be remembered that lipid bilayers behave like soap-bubbles fusing when meeting each other. In this respect interfering proteins should be considered as preventing undesirable and unnecessary fusion and eventually directing the biological membrane fusion process (when, where, how, and overcoming the activation energy). In this latest volume in the renowned Subcellular Biochemistry series, some aspects of fusion of biological membranes as well as related problems are presented. Although not complete, there is a lot of recent information including on virus-induced membrane fusion. The contributors of the chapters are all among the researchers who performed many of the pioneering studies in the field.

Dramatic advances have been made in recent years in the field of redox enzymology which has resulted in an increase of research activities. This volume will cover the recent milestone developments in this field by leading experts, uniting theory and experiment, and selecting contributions to illustrate important aspects of the mechanisms of electron and radical transfer in proteins. Features: A demonstration of the key principles controlling biological redox reactions; Experimental studies of `simple' soluble systems in various enzyme familiies to illustrate concepts in the control of electron transfer reactions; Detail of advances made in membrane electron transfer through structural descriptions of key membrane-embedded proteins; Appeal to those interested in the design and use of redox enzymes, from academics to industrialists.

The last few years have witnessed an explosion of both interest and knowledge about apoptosis, the process by which a cell actively commits suicide. The number of publications on the topic has increased from nothing in the early 1980s to more than 10,000 papers annually today. It is now well recognized that apoptosis is essential in many aspects of normal development and is required for maintaining tissue homeostasis. The idea that life requires death seems somewhat paradoxical, but cell suicide is essential for an animal to survive. For example, without selective destruction of “non-self” T cells, an animal would lack immunity. Similarly, meaningful neural connections in the brain are whittled from a mass of cells. Further, developmental cell remodeling during tissue maturation involves programmed cell death as the major mechanism for functional and structural safe transition of undifferentiated cells to more specialized counterparts. Apoptosis research, with roots in biochemistry, developmental and cell biology, genetics, and immunology, embraces this long-ignored natural law. Failure to properly regulate apoptosis can have catastrophic consequences. Cancer and many diseases (AIDS, Alzheimer’s disease, Parkinson’s disease, heart attack, stroke, etc. ) are thought to arise from deregulation of apoptosis. As apoptosis emerges as a key biological regulatory mechanism, it has become harder and harder to keep up with new developments in this field.

The fluid-mosaic model of membrane structure formulated by Singer and Nicolson in the early 1970s has proven to be a durable concept in terms of the principles governing the organization of the constituent lipids and proteins. During the past 30 or so years a great deal of information has accumulated on the composition of various cell membranes and how this is related to the dif­ ferent functions that membranes perform. Nevertheless, the task of explaining particular functions at the molecular level has been hampered by lack of struc­ tural detail at the atomic level. The reason for this is primarily the difficulty of crystallizing membrane proteins which require strategies that differ from those used to crystallize soluble proteins. The unique exception is bacteriorhodopsin of the purple membrane of Halobacterium halobium which is interpolated into a membrane that is neither fluid nor in a mosaic configuration. To date only 50 or so membrane proteins have been characterised to atomic resolution by diffraction methods, in contrast to the vast data accumulated on soluble proteins. Another factor that has been difficult to explain is the reason why the lipid compliment of membranes is often extremely complex. Many hundreds of different molecular species of lipid can be identified in some membranes. Remarkably, the particular composition of each membrane appears to be main­ tained within relatively narrow limits and its identity distinguished from other morphologically-distinct membranes.

The Coronin Family of Proteins Christoph S. Clemen,* Vasily Rybakin and Ludwig Eichinger he coronins, first described in Dictyostelium discoideum in 1991, have meanwhile been detected in all eukaryotes except plants. They belong to the superfamily of WD40-repeat Tproteins and represent a large family of proteins, which are often involved in cytoskeletal functions. Phylogenetic studies clearly distinguish 12 subfamilies of which six exclusively occur in vertebrates. In the present book we have made a sincere attempt to provide a comprehensive overview on all aspects of coronin proteins including history, structure, subcellular localization and function in different organisms. In addition, we also included a general overview on the WD40 family of proteins and the structurally related Kelch family. The book should be of interest for scientists outside the field, but is more importantly intended as a fast and competent guide for newcomers as well as doctoral and postdoctoral scientists to coronin research in all its facets. The book is divided into four major sections.It provides in the first part an introduction into two superfamilies of proteins with p-propellers, the WD40- and the Kelch-family. Lynn Cooley and Andrew M. Hudson provide evidence that the WD40- and Kelch-repeat families most likely did Figure 1. Condensed phylogenetic tree of the coronin protein family. The tree constitutes the basis of a new nomenclature and shows the evolutionary relationship of the twelve coronin subfamilies {CRN1-CRN12). See also chapter 11-2 by Reginald O. Morgan and M. Pilar Fernandez.

To understand Alzheimer's disease (AD) is one of the major thrusts of present-day clinical research, strongly supported by more fimdamental cellular, biochemical, immunological and structural studies. It is these latter that receive attention within this book. This compilation of 20 chapters indicates the diversity of work currently in progress and summarizes the current state of knowledge. Experienced authors who are scientifically active in their fields of study have been selected as contributors to this book, in an attempt to present a reasonably complete survey of the field. Inevitably, some exciting topics for one reason or another have not been included, for which we can only apologize. Standardization of terminology is often a problem in science, not least in the Alzheimer field; editorial effort has been made to achieve standardization between the Chapters, but some minor yet acceptable personal / author variation is still present, i. e. P-amyloid/amyloid-P; Ap42/Apl-42/APi. 42! The book commences with a broad survey of the contribution that the range of available microscopical techniques has made to the study of Alzheimer's amyloid plaques and amyloid fibrillogenesis. This chapter also serves as an Introduction to the book, since several of the topics introduced here are expanded upon in later chapters. Also, it is significant to the presence of this chapter that the initial discovery of brain plaques, by Alois Alzheimer, utilized light microscopy, a technique that continues to be extremely valuable in present-day AD research.

From being to becoming important, myo-inositol and its derivatives including phosphoinositides and phosphoinositols involved in diversi?ed functions in wide varieties of cells overcoming its insigni?cant role had to wait more than a century. Myo-inositol, infact, is the oldest known inositol and it was isolated from muscle as early as 1850 and phytin (Inositol hexakis phosphate) from plants by Pfeffer in 1872. Since then, interest in inositols and their derivatives varied as the methodology of isolation and puri?cation of the stereoisomers of inositol and their derivatives advanced. Phosphoinositides were ?rst isolated from brain in 1949 by Folch and their structure was established in 1961 by Ballou and his coworkers. After the compilation of scattered publications on cyclitols by Posternak (1965), proceedings of the conference on cyclitols and phosphoinositides under the supervision of Hoffmann-Ostenhof, were p- lished in 1969. Similar proceedings of the second conference on the same s- ject edited by Wells and Eisenberg Jr was published in 1978.In that meeting at the concluding session Hawthorne remarked "persued deeply enough p- haps even myoinositol could be mirror to the whole universe". This is now infact the scenario on the research on inositol and their phosphoderivatives. Finally a comprehensive information covering the aspects of chemistry, b- chemistry and physiology of inositols and their phosphoderivatives in a book entitled Inositol Phosphates written by Cosgrove (1980) was available.

All of the parasitic organisms highlighted in this new book represent medically important human pathogens that contribute significantly to the global burden of disease. As such there is intense interest in understanding the molecular basis of infection by these pathogens—not only with regard to their clinical relevance but also the fascinating biology they reveal. For most of the parasites discussed here the ability to penetrate biological barriers and/or to establish intracellular residence is critical to survival of the pathogen in the mammalian hosts. For other parasites, a tissue invasive phenotype is a key virulence determinant. In the ensuing 18 chapters, select members of this diverse set of protozoan parasites, as well as some examples of the extremely reduced fungal parasites classified as Microsporidia, are discussed within the context of the fascinating molecular strategies employed by these organisms to migrate across biological barriers and to establish residence within target host cells.

Toxigenic Fusarium species and ochratoxigenic fungi are responsible for plant diseases which have important effects on both human and animal health. The development of rapid, robust and sensitive detection methods based on molecular technologies is needed to identify fungal contamination in the field and quantify toxin accumulation in food products. Most of the contributions are from results obtained through the EU 5th Framework project (QLK1-1999-01380) "DETOX-FUNGI: early detection of toxigenic Fusarium species and ochratoxigenic fungi in plant products". Valuable contributions from other scientists have guaranteed a complete overview of increasingly important topic. The present issue contains contributions which cover many aspects of the molecular diversity, phylogeny, variability and PCR-detection of toxigenic fungi and their toxins. Notable amongst these are a review of the phylogeny and diagnosis of Fusarium species and a new system for identifying Fusarium species based on a DNA sequence database. Other papers describe the diversity and analysis of ochratoxigenic Aspergilli and Penicillia.This information will greatly aid those concerned with the early detection of these fungi and their mycotoxins, information which is so essential for overcoming the health risks associated with mycotoxin-contaminated food products.

Many important pharmaceutical drugs, chemicals for the organic synthesis of chiral compounds, electronic elements such as liquid crystals, and polymeric materials are chiral (enantiomeric). The development of new practical methods for the preparation of enantiomerically pure substances is thus vital. Enantiomer Separation is written by several experts working in modern enantiomer separation chemistry who understand the needs of the many scientific and engineering chemists who need a cost-efficient supply of optically active materials of high quality. This book contains the following modern practical methods of enantiomer separation: Practical methods of enantiomer separation are important both in the research laboratory and in industry, especially in the pharmaceutical, fine chemical and electronic industries. Chemists and engineers, as well as students who are working in the field of chiral compounds in universities, institute and industry, will find this book an invaluable resource.

Research is fast paced and advances in RNA interference have recently opened up new opportunities for genetic experiments in human cell lines. However, the possibility to easily modify the genome still remains a powerful tool to investigate the function of coding and regulatory sequences in the vertebrate genome. DT40 has never been a quick and easy road to fame. It would be unfair to blame this on DT40 as it has proven to be a reliable and robust companion with fast doubling time, easy clonability and a relatively stable karyotype. If this model system is going to flourish over the next 15 years, it will be thanks to ingenious and original researchers. They may feel as if they work outside the mainstream, but they can take heart by the fact that only the clever exploitation of diversity and conservation makes biological research both elegant and rewarding. It is with this in mind that the DT40 handbook has been conceived and written. This book provides an up to date overview of the different facets of research, and also intends to help newcomers get started and avoid looming pitfalls. The collection of protocols which have been kindly provided by a number of laboratories will be particularly useful in this regard.

It is more evident now than ever before that dynamic organization of human genome into nucleoprotein structure, chromatin confers the unique regulatory mechanisms for most of the cellular phenomena, which include replication, transcription, DNA repair, recombination and also apoptosis. The dynamic nature of the chromatin is regulated by chromatin modifications (epigenetic alterations), remodeling, histone chaperones and functional interactions of different chromatin interacting n- histone proteins. Dysfunction of this highly inter connected machineries disturb the cellular homoeostasis, and thereby causes several diseases. As we advance in our knowledge of chromatin function and also disease mechanisms in more details, their causal relationship is becoming more evident. This has lead to the identification of chromatin function as target for new generation therapeutics. In the light of these advances, it happens to be the right time to explore current insights into various aspect of chromatin and disease connection under one cover. Authors who are actively involved in chromatin research and have made several original contributions to develop latest paradigms in the field have written the chapters of this book. Significantly, the authors’ repertoire is truly international. They come from eight different countries of Asia, Europe and America. The book has been divided into three different parts. Part I introduces the reader to the dynamic nature of chromatin structure and its link to diseases. First two chapters in this part deal with the chromatin architecture, chromatin dynamics in the cell cycle and molecular mechanism of chromatin remodeling.