It is increasingly clear that signal transduction is a highly organized and integrated process. Divided into two convenient sections, Signal Transduction Protocols, Third Edition focuses on experimental approaches to better understand the complexity of signal transduction. Introductory chapters provide perspective on several of the challenges involved in signal transduction research and offer guidance on selecting the best approaches to various types of questions. The individual chapters provide detailed experimental protocols covering various topics, from the effects of ligand binding on receptor conformation and effector coupling, to moving inside the cell in order to capture the spatial and temporal characteristics of signaling events. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Signal Transduction Protocols serves as an ideal guide to scientists of all backgrounds and is a valuable resource for future progress in the field of signal transduction research.

Membrane Proteins - Engineering, Purification and Crystallization, a volume of Methods In Enzymology, encompasses chapters from the leading experts in the area of membrane protein biology. The chapters provide a brief overview of the topics covered and also outline step-by-step protocol for the interested audience. Illustrations and case example images are included wherever appropriate to help the readers understand the schematics and general experimental outlines.

This book gives an introduction to molecular biophysics. It starts from material properties at equilibrium related to polymers, dielectrics and membranes. Electronic spectra are developed for the understanding of elementary dynamic processes in photosynthesis including proton transfer and dynamics of molecular motors. Since the molecular structures of functional groups of bio-systems were resolved, it has become feasible to develop a theory based on the quantum theory and statistical physics with emphasis on the specifics of the high complexity of bio-systems. This introduction to molecular aspects of the field focuses on solvable models. Elementary biological processes provide as special challenge the presence of partial disorder in the structure which does not destroy the basic reproducibility of the processes. Apparently the elementary molecular processes are organized in a way to optimize the efficiency. Learning from nature by means exploring the relation between structure and function may even help to build better artificial solar cells. The reader is exposed to basic concepts in modern biophysics, such as entropic forces, phase separation, potential of mean force, electron and proton transfer, heterogeneous reactions, coherent and incoherent energy transfer as well as molecular motors. Basic knowledge in classical and Quantum mechanics, electrostatics and statistical physics is desirable. Simplified models are presented which can be solved in limited cases analytically from the guiding lines to generate the basis for a fundamental understanding of the more complex biophysical systems. Chapters close with challenging problems whose solutions are provided at the end of the book to complete the pedagogical treatment in the book. To the second edition several new chapters were added. The medium polarization is treated self-consistently using basic elements of polaron theory and more advanced nonlinear Schroedinger equations to describe the dynamics of solvation. Ion transport through a membrane was extended by the discussion of cooperative effects. Intramolecular transitions are now discussed in the new edition in much more detail, including also radiationless transitions. Very recent developments in spectroscopy are included, especially two-dimensional and hole-burning spectroscopy. The discussion of charge transfer processes was extended by including recent results of hole transfer in DNA in connection with the super-exchange mechanism. The chapter on molecular motors was rewritten to include the most recent developments of new models. The book is a useful text for students and researchers wanting to go through the mathematical derivations in the theories presented. This book attracts a group of applied mathematically oriented students and scholars to the exciting field of molecular biophysics.

How and why certain proteins misfold and how this misfolding is linked to many disease processes has become a well-documented topic of study. Protein Misfolding and Cellular Stress in Disease and Aging: Concepts and Protocols moves beyond the basics to emphasize the molecular effects of protein misfolding at a cellular level, to delineate the impacts and cellular reactions that play a role in pathogenetic mechanisms, and to pinpoint possible manipulations and treatment strategies that can counteract, modify, or delay the consequences of misfolding. The volume begins with several concepts and approaches developed in the recent past including a connection to the research field of aging, where protein misfolding diseases have been equated to premature aging processes, and the book's coverage continues with detailed descriptions of protocols for relevant experimental approaches. Written in the highly successful Methods in Molecular Biology (TM) series format, protocols chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Protein Misfolding and Cellular Stress in Disease and Aging: Concepts and Protocols aims to aid researchers in the field, as well as medical professionals and molecular biologists, in shaping and performing research related to this intriguing and vital subject.

Although less common than -amino acids, non- -amino acids-where the amino group is not on the carbon immediately adjacent to the carboxyl group but is attached to another carbon in the chain (for example, the , , carbon)-are components of biologically important molecules, are significant in the pharmaceutical industry, and are useful starting materials for many areas of organic chemistry. Since the publication of the first edition of this book nearly 20 years ago, synthetic work devoted to the preparation of non- -amino acids has expanded greatly. Methods of Non- -Amino Acid Synthesis, Second Edition has been extensively rewritten and reorganized, providing an up-to-date review of strategies and methods for non- -amino acid synthesis, particularly those amino acids that are key synthetic intermediates or important compounds in their own right. It focuses on acyclic amino acids of C3-C10, but also aminoalkanoic carboxylic acids, aminoalkenoic acids, and aminoalkynoic acids. The new edition contains many updated references and has a greater emphasis on the biological importance of non- -amino acids. In addition to an array of synthetic methods, the book offers discussions on why non- -amino acids are important. The book covers synthetic methods that rely on substituent refunctionalization, the conversion of cyclic precursors to acyclic amino acids, conjugate addition reactions, and enolate anion reactions and condensation reactions that lead to non- -amino acids. It also examines reactions and strategies that lead to good diastereoselectivity and enantioselectivity during synthesis. A chapter devoted to biologically important amino acids includes separate sections on GABA, GABOB, carnitine, DAVA, statine, and other significant amino acids as well as a new section on peptides and proteins that contain non- -amino acids. The final chapter addresses aminocyclic and heterocyclic amino acids.

This book provides a coherent, clear, and uniform presentation of structural, genetic, molecular, and biochemical information available for the zona pellucida domain protein family, which impact pathologies such as infertility, deafness, and cancer. Furthermore it: * Details information about the structure and function of the ZP domain in ZPDC-proteins * Provides illustrations of the organization of ZPDC-proteins, the genes that encode the proteins, and examples of mutations in the ZP domain that cause diseases * Speculates as to the evolution of the ZP domain and potential therapeutics for diseases stemming from ZP domain mutations * Addresses mammalian and non-mammalian systems

Written by a pioneer in the development of spin labeling in biophysics, this expert book covers the fundamentals of nitroxide spin labeling through cutting-edge applications in chemistry, physics, materials science, molecular biology, and biomedicine. Nitroxides have earned their place as one of the most popular organic paramagnets due to their suitability as inhibitors of oxidative processes, as a means to polarize magnetic nuclei, and, in molecular biology, as probes and labels to understand molecular structures and dynamics AS DRAGS FOR CANCER AND OTHER DISEASES. Beginning with an overview of the basic methodology and nitroxides' 145-year history, this book equips students with necessary background and techniques to undertake original research and industry work in this growing field.

This book is devoted to the physical and mathematical modeling of the formation of complexes of protein molecules. The models developed show remarkable sensitivity to the amino acid sequences of proteins, which facilitates experimental studies and allows one to reduce the associated costs by reducing the number of measurements required according to the developed criteria. These models make it possible to reach a conclusion about the interactions between different amino acid chains and to identify more stable sites on proteins. The models also take the phosphorylation of amino acid residues into account. At the end of the book, the authors present possible directions of application of their physical and mathematical models in clinical medicine.

Microbial infection is increasingly seen as a problem as we begin to run out of antibiotics. Understanding how microbes cause disease is essential. In recent years it has begun to emerge that bacteria, fungi, protozoa and viruses can use their cell stress proteins to cause infection. This volume brings together the world's leading experts in the study of the microbial and human cell stress proteins that are involved in enabling microorganisms to infect humans and cause serious disease.

This book focusses on evolutionary, structural and functional aspects of pore-forming proteins, bringing together prominent researchers in the fields of structural biology and cellular and biophysical techniques. The focus is on the MACPF/CDC protein super family that was originally discovered because of unexpected structural similarity between a domain present in bacterial cholesterol-dependent cytolysins (CDC) and proteins of the membrane attack complex/perforin (MACPF) family. Members of the MACPF/CDC super family are crucial for many biological processes, being efficient agents of development, defence, attack and invasion of cells and tissues. However, their best-known role is in bacterial pathogenesis and the proper functioning of the vertebrate immune system, via formation of transmembrane pores in target cell membranes.

The book contains chapters on the distribution of MACPF/CDC proteins and on aspects of their evolution and structural properties, the similarities between different super family members and functional properties of some of the best known examples. The book also contains an overview of biophysical approaches that may be used in the future to provide further insights into how these interesting proteins function.

This detailed book explores the technical breakthroughs with biophysical and cell biological approaches that have advanced the study of molecular mechanisms underlying the generation and transduction of the hedgehog (HH) signal, indicating the important role of sterols in this pathway. Within its pages, the volume examines techniques involving several key protein components in this pathway, including HH, the signaling ligand; Dispatched, a HH secretion regulator; Patched, the receptor of HH; Smoothened, the signal transducer used to transduce the HH signal across the plasma membrane; and GLI, the transcription factor to turn on HH target genes. Written in the highly successful Methods in Molecular Biology series format, chapters feature introductions to their respective topics, lists of the necessary materials and reagents, step-by-step readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Hedgehog Signaling: Methods and Protocols provides researchers in clinical and basic science with a fresh and reliable reference for analyzing HH signal transduction through cell biological, biochemical, and biophysical approaches.

How Proteins Work is an up-to-date and authoritative account of protein function in living systems, explained within the governing parameters of physics, chemistry, and evolution. This text will enable advanced undergraduate students in biochemistry and biophysics to understand the relationships among protein function, structure, and dynamics. It will also serve as a valuable resource for graduate students and researchers looking for a reference on the fundamentals underlying protein function. By providing an integrated view of proteins at both a cellular and systemic level, this textbook shows how evolution drives proteins to adopt domain structures that combine to achieve biological outcomes. The association of proteins into dimers, molecular machines, and multi-enzymatic complexes enables them to achieve catalytic and functional efficiency.

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.

The existence and functioning of intrinsically disordered proteins (IDPs) challenge the classical structure-function paradigm that equates function with a well-defined 3D structure. Uncovering the disordered complement of proteomes and understanding their functioning can extend the structure-function paradigm to herald new breakthroughs in drug development. Structure and Function of Intrinsically Disordered Proteins thoroughly covers the history up to the latest developments in this field.

After examining the principles of protein structure, the classical paradigm, and the history of structural disorder, the book focuses on physical techniques for the identification and characterization of IDPs. It discusses proteomic and bioinformatic approaches and shows how IDPs behave under crowding conditions in living cells. The next several chapters describe the structure, correlating biological processes, and molecular mechanisms of IDPs. The author also explores the evolutionary advancement of structural disorder in proteomes and possible ways of extending the structure-function paradigm to encompass both ordered and disordered states of proteins. He concludes with discussions on the involvement of IDPs in various diseases and how to establish rational drug design through detailed characterization of IDPs.

Although drug discovery rates have leveled off, new insight generated by the study of IDPs may offer fresh strategies for drug development. This work illustrates how these proteins defy the structure-function paradigm and play important regulatory and signaling roles.

In this, the post-genomic age, our knowledge of biological systems continues to expand and progress. As the research becomes more focused, so too does the data. Genomic research progresses to proteomics and brings us to a deeper understanding of the behavior and function of protein clusters. And now proteomics gives way to neuroproteomics as we begin to unravel the complex mysteries of neurological diseases that less than a generation ago seemed opaque to our inquiries, if not altogether intractable. Edited by Dr. Oscar Alzate, Neuroproteomics is the newest volume in the CRC Press Frontiers of Neuroscience Series. With an extensive background in mathematics and physics, Dr. Alzate exemplifies the newest generation of biological systems researchers. He organizes research and data contributed from all across the world to present an overview of neuroproteomics that is practical and progressive. Bolstered by each new discovery, researchers employing multiple methods of inquiry gain a deeper understanding of the key biological problems related to brain function, brain structure, and the complexity of the nervous system. This in turn is leading to new understanding about diseases of neurological deficit such as Parkinson's and Alzheimer's. Approaches discussed in the book include mass spectrometry, electrophoresis, chromatography, surface plasmon resonance, protein arrays, immunoblotting, computational proteomics, and molecular imaging. Writing about their own work, leading researchers detail the principles, approaches, and difficulties of the various techniques, demonstrating the questions that neuroproteomics can answer and those it raises. New challenges wait, not the least of which is the identification of potential methods to regulate the structures and functions of key protein interaction networks. Ultimately, those building on the foundation presented here will advance

Cytochromes P450 (CYPs) comprise a large superfamily of proteins that are of central importance in the detoxification or activation of a tremendous number of natural and synthetic hydrophobic xenobiotics, including many therapeutic drugs, chemical carcinogens and environmental pollutants. CYPs are important in mediating interactions between an organism and its chemical environment and in the regulation of physiological processes. Cytochrome P450 Protocols, Third Edition focuses on high-throughput methods for the simultaneous analysis of multiple CYPs, substrates or ligands. Although the emphasis is on CYPs of mammalian origin, it reflects an increasing interest in CYPs of bacterial species. Also included are chapters on cytochrome P450 reductase (the redox partner of CYPs) and the flavin-containing monooxygenases (FMOs), and metabolomic and lipidomic approaches for identification of endogenous substrates of CYPs ('de-orphanizing' CYP substrates). 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, Cytochrome P450 Protocols, Third Edition provides a wide range of techniques accessible to researchers in fields as diverse as biochemistry, molecular biology, pharmacology, toxicology, environmental biology and genetics.

This volume presents relevant background information to understanding the molecular basis governing unconventional protein secretion (UPS), and in particular explores the latest techniques and protocols that have been successfully applied for the study of this topic. Detailed chapters include an overview of conventional and unconventional secretory pathways along with multidisciplinary approaches and methods used for UPS analysis in different organisms. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Unconventional Protein Secretion: Methods and Protocols will be useful for all interested in the secretory pathway field as well as applications in cell biology, cell development, biomedical research, and healthcare.

Tetraspanin proteins have recently emerged as a new class of modulators of various processes involving cell surface receptors, including cell migration and invasion, host immune responses, cell-cell fusion, and viral infection. The book summarises recent advances in the fields of biology in which the role of tetraspanins have been established and also covers the molecular evolution of the tetraspanin superfamily and structural aspects of the organisation of tetraspanin microdomains.

Upstream processing refers to the production of proteins by cells genetically engineered to contain the human gene which will express the protein of interest. The demand for large quantities of specific proteins is increasing the pressure to boost cell culture productivity, and optimizing bioreactor output has become a primary concern for most pharmaceutical companies. Each chapter in Cell Culture and Upstream Processing is taken from presentations at the highly acclaimed IBC conferences as well as meetings of the European Society for Animal Cell Technology (ESACT) and Protein Expression in Animal Cells (PEACe) and describes how to improve yield and optimize the cell culture production process for biopharmaceuticals, by focusing on safety, quality, economics and operability and productivity issues. Cell Culture and Upstream Processing will appeal to a wide scientific audience, both professional practitioners of animal cell technology as well as students of biochemical engineering or biotechnology in graduate or high level undergraduate courses at university.

Tea is an important non-alcoholic beverage plant of the world. Cultivation of tea is very important as it earns revenue for the tea growing nations especially the developing countries such as India. Although conventional breeding is well-established and has contributed significantly for varietal improvement of this plant and other Camellia species with ornamental value, yet applications of biotechnology are required to intervene some of the issues where conventional breeding is restricted particularly for woody plants such as tea. It is note-worthy to mention that some amounts of biotechnology works in several facets of tea and its wild species have also been done. In the present book, a state-of-the-art on various aspects of breeding and biotechnology has been complied in eight chapters. They are: i) Origin and descriptions of health benefits as well as morphological classification as first chapter, ii) Breeding and cytogenetics that comprise with various conventional approaches of varietal improvement of tea along with their genetic resources, iii) Micropropagation which deals with in-depth study of clonal propagation, iv) Somatic embryogenesis along with alternative techniques such as suspension culture, cry-preservation etc. v) Molecular breeding that deals with application of various DNA-based markers, linkage map etc., vi) Genetic transformation and associated factors, vii) Stress physiology complied with various works done in tea along with its wild relatives on abiotic as well as biotic stress, and viii) Functional genomics that describe the various works of molecular cloning and characterizations, differential gene expression, high-throughput sequencing, bioinformatics etc. Importantly, the author has made exclusive tables in most of the chapters that include the summary of the works in particular topic. In a nutshell, the book compiles the work already been done, identifies the problems, analyzes the gaps on breeding and biotechnological works of tea as well as its wild species and discusses the future scope as conclusion. Every effort has been made to include all the published works till June 2013. The book will be a useful resource for post-graduate, doctoral as well post-doctoral students working on tea as well as other woody plants. This will also be useful for the scientists working in the areas of life sciences, genomics, biotechnology and molecular biology.

Site-specific mutagenesis of DNA, developed some thirty years ago, has proven to be one of the most important advances in biology. By allowing the site-specific replacement of any amino acid in a protein with one of the other nineteen amino acids, it ushered in the new era of "Protein Engineering." The field of protein engineering has, however, evolved rapidly since then and the last fifteen years have witnessed remarkable advances through the use of new chemical, biochemical and molecular biological tools towards the synthesis and manipulation of proteins. The chapters included in this book reflect the rapid evolution of protein engineering and its many applications in basic research, biotechnology, material sciences and therapy. This book will provide the reader with an introduction to state-of the-art concepts and methods and will be of use to anyone interested in the study of proteins, in academia as well as in industry.

This book is specifically about the application of the extremely powerful interaction between the protein avidin or its homologues and the vitamin biotin and some of its homologues. With excellent descriptions of laboratory protocols written by expert researchers, this volume is equally perfect for the student or the professional laboratory scientist.

This book surveys the current knowledge concerning the expression and function of small stress proteins (sHsps) in different organisms, ranging from prokaryotes to humans. It provides an overview of the diversity and complex evolutionary history of sHsps and describes their function and expression in different eukaryote models. Additional chapters discuss the involvement of sHsps in pathological conditions and gene therapy approaches towards a control of sHsp expression levels.

This book provides the basis for understanding the elastic properties of nucleic acids (DNA, RNA), the methods used to manipulate them (e.g. optical, magnetic and acoustic tweezers and traps), and how to observe their interactions with proteins (e.g. fluorescence microscopy, FCS, FRET, etc.). It then exemplifies the use of these various methods in the study of three families of DNA enzymes: polymerases, helicases and topoisomerases. The book aims not to be exhaustive, but rather to stimulate the imagination of readers in the application of these single molecule approaches to the study of DNA/RNA and their interactions.

This new edition features research from nearly 60 of the profession's most distinguished international authorities. Recognizing emerging developments in biopolymer systems research with fully updated and expanded chapters, the second edition discusses the biopolymer-based multilayer structures and their application in biosensors, the progress made in the understanding of protein behaviour at the air-water interface, experimental findings in ellipsometry and reflectometry, and recent developments concerning protein interfacial behaviour in microfabricated total analysis systems and microarrays. With over 3000 references, this is an essential reference for professionals and students in surface, pharmaceutical, colloid, polymer, and medicinal chemistry; chemical, formulation, and application engineering; and pharmacy.