This detailed volume compiles state-of-the-art protocols that will serve as recipes for scientists researching collagen, an abundant protein with great importance to health and disease, as well as in applications like food, cosmetics, pharmaceuticals, cosmetic surgery, artificial skin, and glue. Beginning with a section on in vitro models for the characterization of collagen formation, the book continues by highlighting large-scale analysis of collagen with mass spectrometry in order to elucidate the proteomics, degradomics, interactomes, and cross-linking of collagen, high resolution imaging approaches for collagen by the use of scanning electron microscopy and multiphoton imaging, as well as the role of collagen during physiological and pathological conditions. Written for the highly successful Methods in Molecular Biology series, 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 practical, Collagen: Methods and Protocols is an ideal guide to high quality and repeatable protocols in this vital field of study.

This volume provides the reader with detailed protocols that describe a variety of in vitro and in vivo techniques that study reductionist systems and human samples. The methods covered in this book explore a broad selection of topics such as functional aspects of alpha-synuclein biology, its lipid-interactions and misfolding, and functional deficits in cells and mice. Chapters also cover topics such as kinetic measurements of endocytosis and exocytosis in cultured neurons; electrochemiluminescence-based detection; yeast-based screens to target alpha-synuclein toxicity; mass-spectrometry of alpha-synuclein in rat cortical neurons; and expression and purification of untagged a-synuclein. 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. Cutting-edge and thorough, Alpha-Synuclein: Methods and Protocols is a valuable reference guide that aids researchers working on topics related to alpha-synuclein and its various aspects.

Cancer is one of the leading death cause of human population increasingly seen in recent times. Plants have been used for medicinal purposes since immemorial times. Though, several synthetic medicines are useful in treating cancer, they are inefficient and unsafe. However, plants have proved to be useful in cancer cure. Moreover, natural compounds from plants and their derivatives are safe and effective in treatment and management of several cancer types. The anticancer plants such as Catharanthus roseus, Podophyllum peltatum, Taxus brevifolia, Camptotheca acuminate, Andrographis paniculata, Crateva nurvala, Croton tonkinensis, Oplopanax horridus etc., are important source of chemotherapeutic compounds. These plants have proven their significance in the treatment of cancer and various other infectious diseases. Nowadays, several well-known anticancer compounds such as taxol, podophyllotoxins, camptothecin, vinblastine, vincristine, homoharringtonine etc. have been isolated and purified from these medicinal plants. Many of them are used effectively to combat cancer and other related diseases. The herbal medicine and their products are the most suitable and safe to be used as an alternative medicine. Based on their traditional uses and experimental evidences, the anticancer products or compounds are isolated or extracted from the medicinally important plants. Many of these anticancer plants have become endangered due to ruthless harvesting in nature. Hence, there is a need to conserve these species and to propagate them in large scale using plant tissue culture. Alternatively, plant cell tissue and organ culture biotechnology can be adopted to produce these anticancer compounds without cultivation. The proper knowledge and exploration of these isolated molecules or products could provide an alternative source to reduce cancer risk, anti-tumorigenic properties, and suppression of carcinogen activities. Anticancer plants: Volume 1, Properties and Application is a very timely effort in this direction. Discussing the various types of anticancer plants as a source of curative agent, their pharmacological and neutraceutical properties, cryo-preservations and recent trends to understand the basic cause and consequences involved in the diseases diagnosis. We acknowledge the publisher, Springer for their continuous inspiration and valuable suggestions to improvise the content of this book. We further extend our heartfelt gratitude to all our book contributors for their support, and assistance to complete this assignment. I am sure that these books will benefit the scientific communities including academics, pharmaceuticals, nutraceuticals and medical practitioners.

Using both epidemiological and model organism approaches, we have gained new insights into the physiological and molecular aspects of aging, which has led to significant advancements in potential anti-aging strategies. Reviews on Biomarker Studies in Aging and Anti-Aging Research presents a series of reviews in various aspects of aging and age-related disease research along with several methods which have shown progress as potential anti-aging approaches. The book is aimed at researchers in the areas of aging and chronic disease, as well as to clinical scientists, physicians and major drug companies. It provides important information on disease mechanisms, and each chapter is presented in the context of the aging process, specific chronic diseases or different therapeutic areas.

This book is the first to be entirely devoted to the challenging art of handling membrane proteins out of their natural environment, a key process in biological and pharmaceutical research, but one plagued with difficulties and pitfalls. Written by one of the foremost experts in the field, Membrane Proteins in Aqueous Solutions is accessible to any member of a membrane biology laboratory. After presenting the structure, functions, dynamics, synthesis, natural environment and lipid interactions of membrane proteins, the author discusses the principles of extracting them with detergents, the mechanisms of detergent-induced destabilization, countermeasures, and recent progress in developing detergents with weaker denaturing properties. Non-conventional alternatives to detergents, including bicelles, nanodiscs, amphipathic peptides, fluorinated surfactants and amphipols, are described, and their relative advantages and drawbacks are compared. The synthesis and solution properties of the various types of amphipols are presented, as well as the formation and properties of membrane protein/amphipol complexes and the transfer of amphipol-trapped proteins to detergents, nanodiscs, lipidic mesophases, or living cells. The final chapters of the book deal with applications: membrane protein in vitro folding and cell-free expression, solution studies, NMR, crystallography, electron microscopy, mass spectrometry, amphipol-mediated immobilization of membrane proteins, and biomedical applications. Important features of the book include introductory sections describing foundations as well as the state-of-the-art for each of the biophysical techniques discussed, and topical tables which organize a widely dispersed literature. Boxes and annexes throughout the book explain technical aspects, and twelve detailed experimental protocols, ranging from in vitro folding of membrane proteins to single-particle electron cryomicroscopy, have been contributed by and commented on by experienced users. Membrane Proteins in Aqueous Solutions offers a concise, accessible introduction to membrane protein biochemistry and biophysics, as well as comprehensive coverage of the properties and uses of conventional and non-conventional surfactants. It will be useful both in basic and applied research laboratories and as a teaching aid for students, instructors, researchers, and professionals within the field.

In this book, the author describes the development of the experimental diffraction setup and structural analysis of non-crystalline particles from material science and biology. Recent advances in X-ray free electron laser (XFEL)-coherent X-ray diffraction imaging (CXDI) experiments allow for the structural analysis of non-crystalline particles to a resolution of 7 nm, and to a resolution of 20 nm for biological materials. Now XFEL-CXDI marks the dawn of a new era in structural analys of non-crystalline particles with dimensions larger than 100 nm, which was quite impossible in the 20th century. To conduct CXDI experiments in both synchrotron and XFEL facilities, the author has developed apparatuses, named KOTOBUKI-1 and TAKASAGO-6 for cryogenic diffraction experiments on frozen-hydrated non-crystalline particles at around 66 K. At the synchrotron facility, cryogenic diffraction experiments dramatically reduce radiation damage of specimen particles and allow tomography CXDI experiments. In addition, in XFEL experiments, non-crystalline particles scattered on thin support membranes and flash-cooled can be used to efficiently increase the rate of XFEL pulses. The rate, which depends on the number density of scattered particles and the size of X-ray beams, is currently 20-90%, probably the world record in XFEL-CXDI experiments. The experiment setups and results are introduced in this book. The author has also developed software suitable for efficiently processing of diffraction patterns and retrieving electron density maps of specimen particles based on the diffraction theory used in CXDI.

After decades of dominance of genetics and genomics, the importance of structural biology is growing exponentially in the field of plant biology. The main objectives of this new book series is to "demystify" structural biology for plant researchers and to provide important insights into the basic molecular mechanisms underlying plant development through the diverse approaches utilized by structural biologists. The book series starts with a theme dedicated to hormonal signaling that has benefited from the application of structural biology. "Plant Structural Biology: Hormonal Regulations" provides up-to-date knowledge of the structural aspects of hormonal signal recognition, signal transduction, hormonal control of downstream regulatory pathways and hormonal crosstalk. The most distinctive features of this book as well as future titles is/will be to provide overview of cutting-edge research in the field of plant structural biology, and to serve as a compendium of various approaches that could be applied to problems being solved in modern plant biology. Last but not least, we hope this book will facilitate and broaden the community of (not only) plant scientists who are interested in structural biology approaches and tools. For these reasons, the style of this series is concise and general, in order to avoiding unnecessary details. Explanatory boxes describing the basics of specific approaches (e.g. X-ray crystallography, NMR, SAXS, molecular dynamics simulations, etc.) are included.

This volume takes a closer look how the cell organelles Golgi apparatus (also known as the Golgi complex or Golgi body), and centriole are structurally and functionally intertwined. Initially, it was believed that the role of Golgi complex is limited to the packaging and preparation for secretion of various cellular proteins, while the centriole participates in cell division and cilia formation. However, since their discovery nearly 200 years ago, it became clear that these two organelles are interacting, and that their functions are much more complex and far reaching than previously thought. Recent findings indicate that the Golgi-Centriole relationship may be important for directional protein transport, cell polarization and cell cycle progression. Current studies indicate that Golgi and centriole also participate in development and act as cellular and immunological sensors, and that their abnormalities lead to cell and developmental abnormalities, Alzheimer, cancer, various lipid disorders and neurological and immunological diseases in humans. This volume combines the latest information on the structure, molecular composition, and roles of Golgi and centriole in various cellular functions and diseases. The better understanding of the Golgi-centriole interactions may lead to the development of novel therapies for the treatment of various diseases, including cancer.

This book provides a timely state-of-the-art overview of voltage-gated sodium channels, their structure-function, their pharmacology and related diseases. Among the topics discussed are the structural basis of Na+ channel function, methodological advances in the study of Na+ channels, their pathophysiology and drugs and toxins interactions with these channels and their associated channelopathies.

This volume brings together a plethora of protocols and experimental methods used by scientists to study calpains, their inhibitors, and their substrates. It also explores bioinformatic approaches to calpain substrate identification. The chapters in this book are divided into five parts and cover topics such as production and purification of calpains; determination of calpain localization, expression, and activity; identification of calpain-activated protein function; interrogation of calpastatin; and manipulation of calpain expression. 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. Cutting-edge and practical, Calpain: Methods and Protocols is a valuable resource for researchers and scientists who want to learn more about this developing field, and get inspired to make new discoveries that will aid in diagnosing and treating calpain-related diseases.

This book shows the various sandwich assays that are constructed from recognition molecules, such as antibodies, oligonucleotide sequences and aptamers, developed as a result of nano- and biotechnology advances. It consists of ten chapters presenting interesting examples of these assays, organized according to the type of analytic methods (colorimetric, fluorescence, electrochemical, etc.) and detected objects (protein, nucleic acid, small-molecule, ion, etc.). It also includes a chapter discussing the introduction of sandwich assays as biosensors for the detection of a range of targets. It is an interesting and useful resource for a wide readership in various fields of chemical science and nanotechnology.

Experts from around the world review the current field of the immunobiology of heat shock proteins, and provide a comprehensive account of how these molecules are spearheading efforts in the understanding of various pathways of the immune system. This one-stop resource contains numerous images to both help illustrate the research on heat shock proteins, and better clarify the field for the non-expert. Heat shock proteins (HSPs) were discovered in 1962 and were quickly recognized for their role in protecting cells from stress. Twenty years later, the immunogenicity of a select few HSPs was described, and for the past 30 years, these findings have been applied to numerous branches of immunology, including tumor immunology and immunosurveillance, immunotherapy, etiology of autoimmunity, immunotherapy of infectious diseases, and expression of innate receptors. While HSPs can be used to manipulate immune responses by exogenous administration, they appear to be involved in initiation of de novo immune responses to cancer and likely in the maintenance of immune homeostasis.

This book introduces characteristic features of the protein structure prediction (PSP) problem. It focuses on systematic selection and improvement of the most appropriate metaheuristic algorithm to solve the problem based on a fitness landscape analysis, rather than on the nature of the problem, which was the focus of methodologies in the past. Protein structure prediction is concerned with the question of how to determine the three-dimensional structure of a protein from its primary sequence. Recently a number of successful metaheuristic algorithms have been developed to determine the native structure, which plays an important role in medicine, drug design, and disease prediction. This interdisciplinary book consolidates the concepts most relevant to protein structure prediction (PSP) through global non-convex optimization. It is intended for graduate students from fields such as computer science, engineering, bioinformatics and as a reference for researchers and practitioners.

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.

This new edited volume in the Springer Subcellular Biochemistry Series presents a comprehensive, state-of-the-art overview of the proteomics of peroxisomes derived from mammalian, Drosophila, fungal, and plant origin, and contains contributions from leading experts in the field. The development of sensitive proteomics and mass spectrometry technologies, combined with bioinformatics approaches now allow the identification of low-abundance and transient peroxisomal proteins and permits to identify the complete proteome of peroxisomes, with the consequent increase of our knowledge of the metabolic and regulatory networks of these important cellular organelles. The book lines-up with these developments and is organized in four sections including: (i) mass spectrometry-based organelle proteomics; (ii) prediction of peroxisomal proteomes; (iii) analysis of peroxisome proteome interaction networks; and (iv) peroxisomes in relation to other subcellular compartments. The editor Luis A. del Rio is Professor ad honorem of the Spanish National Research Council (CSIC) in the Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Biochemistry and Cell & Molecular Biology of Plants, at the Estacion Experimental del Zaidin, Granada, Spain. Del Rio's research group focuses on the metabolism of reactive oxygen species (ROS), reactive nitrogen species (RNS) and antioxidants in plant peroxisomes, and the ROS- and RNS-dependent role of peroxisomes in plant cell signalling. The editor Michael Schrader is Professor of Cell Biology & Cytopathology in the Department of Biosciences at the University of Exeter, UK. Using mammalian peroxisomes as model organelles, Prof. Schrader and his team aim to unravel the molecular machinery and signalling pathways that mediate and regulate the formation, dynamics and abundance of these medically relevant cellular compartments.

This book comprehensively reviews the state-of-the-art strategies developed for protein-protein interaction (PPI) inhibitors, and highlights the success stories in new drug discovery and development. Consisting of two parts with twelve chapters, it demonstrates the design strategies and case studies of small molecule PPI inhibitors. The first part discusses various discovery strategies for small molecule PPI inhibitors, such as high throughput screening, hot spot-based design, computational approaches, and fragment-based design. The second part presents recent advances in small molecule inhibitors, focusing on clinical candidates and new PPI targets. This book has broad appeal and is of significant interest to the pharmaceutical science and medicinal chemistry communities.

The Subcellular Biochemistry series has recently embarked upon an almost encyclopaedic coverage of topics relating to the structure and function of macromolecular complexes (Volumes 82, 83 and 87). The present multi-author text covers numerous aspects of current research into molecular virology, with emphasis upon viral protein and nucleoprotein structure and function. Structural data from cryo-electron microscopy and X-ray crystallography is displayed throughout the book. The 17 chapters in the book cover diverse interesting topics, all currently under investigation, contributed by authors who are active actively involved in present-day research. Whilst structural aspects predominate, there is much consideration of the structure-function relationship. In addition, the book correlates with and extends from Volume 68 of the series "Structure and Physics of Viruses: An Integrated Textbook". This book is directed primarily at professionals that work in the broad field of Structural Biology and will be of particular interest to Structural Virologists. The editors, David Bhella and Robin Harris, have much experience in virology and protein structure, respectively. Dr Bhella is Director of the Scottish Macromolecular Imaging Centre. Professor Robin Harris is the long-standing Series Editor of the Subcellular Biochemistry series. He has edited and contributed to several books in the series.

This book provides information about the sources, structure, and properties of keratin as well as its applications. The extraction from different biomass sources (e.g. feathers, hairs, nails, horn, hoof, and claws) as well as the characterization methods of these extracted materials are explained. The development of bioproducts from keratins is challenging and limited since they are neither soluble in polar solvents nor in non-polar solvents. Therefore, the utilization of different microorganisms for the degradation of keratin is also discussed. The main aim of this book is to highlight the unique features of keratin and to update readers with the possible prospects to develop various value-added products from keratins. The book is highly interesting to researchers working in industry and academia on bioproducts, tissue engineering, biocomposites, biofilm, and biofibers.

This volume explores numerous techniques for the genetic, molecular, biochemical, and structural examination of BCL-2 family proteins and their interactions. The chapters in this book cover topics such as the relevance of BCL-2 proteins in health and disease; evaluating cellular dependencies to specific BCL-2 family proteins; flow-cytometry-based methods for measuring BCL-2 proteins and mitochondrial-based cell death; measuring activity and interactions of BCL-2 family proteins in the presence of mitochondria, artificial membranes or yeast; conformational activation and oligomerization of pro-apoptotic proteins BAX and BAK leading to cytochrome c release and apoptosis; structural and biophysical studies in solution and lipid vesicles using nuclear magnetic resonance, cryo-electron microscopy, fluorescence microscopy and electron paramagnetic resonance. 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. Cutting-edge and thorough, BCL-2 Family Proteins: Methods and Protocols is a valuable resource to inspire and encourage novice and established scientists to further their research and make new discoveries in this exciting field.

This book will provide current understandings about two ubiquitously expressed metabotropic GPCRs, G-coupled purinoreceptor type 2 (P2Y) and Takeda G-protein-coupled bile acid receptor 5 (TGR5). G protein coupled receptors (GPCRs) are the largest family of proteins implicated in majority of cellular responses. The two receptor sub-families play a central role in many physiological functions as well as in many pathological conditions. This book offers up-to-date information on the physiological functions, signaling pathways and regulatory mechanisms of P2Y and TGR5 receptors. In addition, this book provides a comprehensive overview about the abnormalities of P2Y/TGR5 receptors and their contribution in the development and progression of pathological conditions. It also covers the currently available natural, chemical and pharmacological agents targeting these two receptor families and their therapeutic implications in P2Y and TGR5 associated disorders. This book is a valuable source for beginners and researchers to follow the rapidly progressing field of these two GPCR subfamily members.

Hemagglutinins refers to glycoproteins which bring about agglutination of erythrocytes or hemagglutination. Hemagglutination can be used to identify surface antigens on erythrocytes (with known antibodies) and, hence, the blood type of an individual. Hemagglutinins consist of lectins and antibodies. Lectins (from the Latin legere, "to select") are non-immune glycoproteins that exhibit reversible binding to specific carbohydrate structures, glycans of glycoproteins, glycolipids and polysaccharides. Lectins own at least one non-catalytic domain, and, in some cases, a few carbohydrate binding domains, which enable them to effect agglutination of erythrocytes and other cells or precipitation of glycoconjugates. Lectins are found in a constellation of organisms and display an array of activities. Topics discussed in this book encompass algal lectins, plant type 2 ribosome inactivating proteins, edible legume lectins, jacalin, jacalin-related lectin, wheat lectins, rice lectins, banana lectins and potato lectin, immunomodulatory action of plant lectins, piezoelectric assay using the galactose-binding Bauhinia monandra leaf lectin and its antibody as a potent tool for detection of antigen-antibody recognition, assays using hemagglutinins of different origins to investigate their mechanisms of action, lectins from medicinal herbs and medicinal mushrooms, C-type lectins from a diversity of invertebrates and vertebrates, fish lectins and amphibian lectins.

This new edited volume in the Springer Subcellular Biochemistry Series presents a comprehensive, state-of-the-art overview of the proteomics of peroxisomes derived from mammalian, Drosophila, fungal, and plant origin, and contains contributions from leading experts in the field. The development of sensitive proteomics and mass spectrometry technologies, combined with bioinformatics approaches now allow the identification of low-abundance and transient peroxisomal proteins and permits to identify the complete proteome of peroxisomes, with the consequent increase of our knowledge of the metabolic and regulatory networks of these important cellular organelles. The book lines-up with these developments and is organized in four sections including: (i) mass spectrometry-based organelle proteomics; (ii) prediction of peroxisomal proteomes; (iii) analysis of peroxisome proteome interaction networks; and (iv) peroxisomes in relation to other subcellular compartments. The editor Luis A. del Rio is Professor ad honorem of the Spanish National Research Council (CSIC) in the Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Biochemistry and Cell & Molecular Biology of Plants, at the Estacion Experimental del Zaidin, Granada, Spain. Del Rio's research group focuses on the metabolism of reactive oxygen species (ROS), reactive nitrogen species (RNS) and antioxidants in plant peroxisomes, and the ROS- and RNS-dependent role of peroxisomes in plant cell signalling. The editor Michael Schrader is Professor of Cell Biology & Cytopathology in the Department of Biosciences at the University of Exeter, UK. Using mammalian peroxisomes as model organelles, Prof. Schrader and his team aim to unravel the molecular machinery and signalling pathways that mediate and regulate the formation, dynamics and abundance of these medically relevant cellular compartments.

This volume provides an up-to-date, in-depth overview of the methods that have been applied to studying the complex metalloproteins at a molecular level. Chapters cover a wide range of approaches focusing on genetic, biochemical, spectroscopic, chemical methods, and theoretical calculations. 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, Metalloproteins: Methods and Protocols aims to be useful for anyone who is interested in metalloprotein research and wants to address the unanswered mechanistic and biosynthetic questions of these fascinating enzyme systems.

This volume describes protocols for basic state-of-the-art approaches in the field of peptidomics. Most of these approaches are independent of the instruments used for analysis and can easily be adapted for equipment that is available in a typical proteomics facility. Chapters detail many of the basic techniques used to detect and identify peptides, methods for the relative quantitation of peptides between samples using isotopic labels or label-free approaches, and biological species as well as sample types. Written in the highly successful format of the Methods in Molecular Biology series, each chapter includes an introduction to the topic, a list of the necessary materials and reagents, reproducible step-by-step laboratory protocols, and tips on troubleshooting common problems and avoiding pitfalls. Authoritative and practical, Peptidomics: Methods and Strategies provides useful guidance for studies in the rapidly growing field of peptidomics.

This book examines Au (I, III) complexes that selectively attack and inhibit zinc finger proteins (ZnFs) for potential therapeutic use. The author explores gold(I)-phosphine, gold(III) complexes with N^N and C^N donors as inhibitors of the HIV-1 nucleocapsid protein (NCp7), in comparison to the human transcription factor Sp1. To determine the coordination sphere of the gold adducts formed by interaction with ZnFs, two innovative approaches are used, based on Travelling-Wave Ion Mobility coupled with Mass Spectrometry (TWIM-MS), and X-ray Absorption Spectroscopy. Both approaches are proven to yield valuable structural information regarding the coordination sphere of gold in the adducts. In addition, the organometallic compound [Au (bnpy)Cl2] is evaluated. The system is shown to be capable of inhibiting ZnFs by means of C-S coupling.