This volume presents an overview of nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) structure and function. It then continues with methods for the analysis of these pathways including conventional enzymological assays, contemporary mass spectrometric analysis techniques, specialized molecular biological approaches applicable to NRPSs and PKSs, and small molecule analysis tools tailored to this very special class of natural products, and concludes by examining bioinformatics tools for the analysis of these enzymes, pathways, and molecules. 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, Nonribosomal Peptide and Polyketide Biosynthesis: Methods and Protocols serves as a valuable reference for those experienced in studying NRPS and PKS enzymes, pathways, and natural products as well as a gateway for those just entering the field.

Lipases are the most applied enzymes in organic synthesis due to their broad substrate acceptance and because of the availability of the molecular, biochemical, themodynamical and solvent engineering tools, which allows the optimization of lipases and lipase-catalyzed reactions. On the other hand, phospholipases are emerging as useful enzymes in food and pharmaceutical industries. In Lipases and Phospholipases: Methods and Protocols, expert researchers in the field provide key techniques to investigate these essential enzymes. Focusing on fundamental issues, current and new applications as well as practical step-by-step protocols, and the extensive applications of lipases and the potential application of phospholipases and its inhibitors. Written in the highly successful Methods in Molecular Biology (TM) series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory. Thorough and intuitive, Lipases and Phospholipases: Methods and Protocols aids scientists in continuing to study lipases, phopholipases and related enzymes.

This contributed volume sheds new light on waste management and the production of biofuels. The authors share insights into microbial applications to meet the challenges of environmental pollution and the ever- growing need for renewable energy. They also explain how healthy and balanced ecosystems can be created and maintained using strategies ranging from oil biodegration and detoxification of azo dyes to biofouling. In addition, the book illustrates how the metabolic abilities of microorganisms can be used in microbial fuel-cell technologies or for the production of biohydrogen. It inspires young researchers and experienced scientists in the field of microbiology to explore the application of green biotechnology for bioremediation and the production of energy, which will be one of the central topics for future generations.

This thesis describes a series of investigations designed to assess the value of metalloenzymes in systems for artificial and adapted photosynthesis. The research presented explores the interplay between inherent enzyme properties such as structure, rates and thermodynamics, and the properties of the semiconducting materials to which the enzyme is attached. Author, Andreas Bachmeier provides a comprehensive introduction to the interdisciplinary field of artificial photosynthesis, allowing the reader to grasp the latest approaches being investigated, from molecular systems to heterogeneous surface catalysis. Bachmeier's work also uses metalloenzymes to highlight the importance of reversible catalysts in removing the burden of poor electrocatalytic rates and efficiencies which are common characteristics for most artificial photosynthesis systems. Overall, this thesis provides newcomers and students in the field with evidence that metalloenzymes can be used to establish new directions in artificial photosynthesis research.

NMR Spectroscopy for Chemical Analysis at Low Magnetic Fields, by Stefan Gloggler, Bernhard Blumich, Stephan Appelt

Dynamic Nuclear Hyperpolarization in Liquids, by Ulrich L. Gunther

NMR with Multiple Receivers, by Eriks Kupce

TROSY NMR Spectroscopy of Large Soluble Proteins, by Yingqi Xu, Stephen Matthews

Solid-State NMR Spectroscopy of Proteins, by Henrik Muller, Manuel Etzkorn, Henrike Heise

Paramagnetic Solid-State Magic-Angle Spinning NMR Spectroscopy, by Guido Pintacuda, Gwendal Kervern"

This book brings together reviews from international experts who are exploring the biological activities of nanomaterials for medical applications or to better understand nanotoxicity. Topics include but are not limited to the following: 1) mechanistic understanding of nanostructure-bioactivity relationships; 2) the regulation of nanoparticles' bioactivity by means of chemical modification; 3) the new methodologies and standard methods used to assess nanoparticles' bioactivity; 4) the mechanisms involved in nanoparticle-biomolecule interactions and nanoparticle-cell interactions; and 5) biomedical applications of nanotechnology. The book will be a valuable resource for a broad readership in various subfields of chemical science, engineering, biology, environment, and medicine.

This volume details the importance of multiple experimental techniques and computational methods needed to obtain the comprehensive picture of protein complex structure, dynamics and assembly afforded by the emerging field of integrative structural biology. Chapters guide readers through the broad spectrum of approaches required for a complete representation of protein complexes, including expression and purification, experimental characterization of structure and assembly, and computational methods for identifying protein complexes and modelling their assembly. 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, Protein Complex Assembly: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Whether the pursuit is commercially motivated or purely academic, engineering a novel biological catalyst is an enticing challenge. High-resolution protein structure analysis allows for rational alteration of enzyme function, yet many useful enzyme variants are the product of well-designed selection schemes or screening strategies. Enzyme Engineering: Methods and Protocols provides guidance to investigators wishing to create enzyme variants with desired properties. This detailed volume covers such topics as a simple method for generating site-specific mutations within bacterial chromosomes. It also highlights the engineering of two difference types of rare-cutting endonucleases that show great potential in gene therapy applications: The newest development is the emergence of TAL effector nucleases or TALENs. Chapters describe newly developed technologies in sufficient detail so that each method can be practiced in a standard molecular biology laboratory. 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 Enzyme Engineering: Methods and Protocols will be valuable for scientists with a budding interest in protein engineering as well as veterans looking for new approaches to apply in established discovery programs.

This volume reviews numerous reaction mechanisms and applications of nucleic acids with catalytic activity. Written by an interdisciplinary team of authors, it provides an essential overview of these acids' fundamental aspects, while also addressing associated methodologies such as nucleic acid enzyme engineering, peroxidase-mimicking DNAzymes and Aptazymes. After the discovery of natural ribozymes - RNA molecules that mediate the cleavage and formation of phosphodiester bonds and the formation of peptide bonds - numerous artificial ribozymes with altered catalytic activities were produced by in vitro and in vivo selection. Unlike ribozymes, DNAzymes do not occur in nature. Although the catalytic activity of nucleic acid enzymes is usually much slower than that of proteins, nucleic acid enzymes with comparable catalytic activity have been obtained using stringent selection processes. The key advantages of these enzymes: they are e.g. smaller, easier to produce and purify than proteins, and can withstand denaturation, e.g. by heat. Over the last few years, the number of publications on the applications of enzymatic nucleic acids has grown steadily. Summarizing the fundamentals and applications of these acids, this book will not only be an excellent resource for experts in the field but will also guide young researchers just starting out in this significant area.

One of the major challenges of modern biology and medicine consists in finding means to visualize biomolecules in their natural environment with the greatest level of accuracy, so as to gain insight into their properties and behaviour in a physiological and pathological setting. This has been achieved thanks to the design of novel imaging agents, in particular to fluorescent biosensors.

Fluorescence Biosensors comprise a large set of tools which are useful for fundamental purposes as well as for applications in biomedicine, drug discovery and biotechnology. These tools have been designed and engineered thanks to the combined efforts of chemists and biologists over the last decade, and developed hand in hand together with imaging technologies.

This volume will convey the many exciting developments the field of fluorescent biosensors and reporters has witnessed over the recent years, from concepts to applications, including chapters on the chemistry of fluorescent probes, on technologies for monitoring protein/protein interactions and technologies for imaging biosensors in cultured cells and in vivo. Other chapters are devoted to specific examples of genetically-encoded reporters, or to protein and peptide biosensors, together with examples illustrating their application to cellular and in vivo imaging, biomedical applications, drug discovery and high throughput screening.

Key features:

* Contributions from leading authorities * Informs and updates on all the latest developments in the field

This new volume of "Methods in Enzymology" continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. This volume coversFluorescence Fluctuation Spectroscopy
Contains chapters on such topics as Time-integrated fluorescence cumulant analysis, Pulsed Interleaved Excitation, and raster image correlation spectroscopy and number and brightness analysis.
Continues the legacy of this premier serial with quality chapters authored by leaders in the fieldCovers fluorescence fluctuation spectroscopyContains chapters on such topics as time-integrated fluorescence cumulant analysis, pulsed interleaved excitation, and raster image correlation spectroscopy and number and brightness analysis."

This new volume of "Methods in Enzymology" continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers fluorescence fluctuation spectroscopy and includes chapters on such topics as Forster resonance energy transfer (fret) with fluctuation algorithms, protein corona on nanoparticles by FCS, and FFS approaches to the study of receptors in live cells.

Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers fluorescence fluctuation spectroscopy Contains chapters on such topics as Forster resonance energy transfer (fret) with fluctuation algorithms, protein corona on nanoparticles by FCS, and FFS approaches to the study of receptors in live cells"

Since the publishing of the first edition, the methodologies and instrumentation involved in the field of mass spectrometry-based proteomics has improved considerably. Fully revised and expanded, Mass Spectrometry Data Analysis in Proteomics, Second Edition presents expert chapters on specific MS-based methods or data analysis strategies in proteomics. The volume covers data analysis topics relevant for quantitative proteomics, post translational modification, HX-MS, glycomics, and data exchange standards, among other topics. Written in the highly successful Methods in Molecular Biology series format, chapters include brief introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Updated and authoritative, Mass Spectrometry Data Analysis in Proteomics, Second Edition serves as a detailed guide for all researchers seeking to further our knowledge in the field of proteomics.

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.

Carbohydrate antigens on glycoconjugates of mammalian cells play crucial roles in various biological processes and are epitopes recognized by the immune system, as glycobiology has hugely been progressed during the past two decades. The book focuses on sialic acid-based xenoantigenes. In pig to human xenotransplantation, exposure of pig organs to human blood results in hyper acute rejection (HAR), caused by differences in carbohydrate epitopes between human and pig vascular endothelia. Although Gal-antigen as major antigen was eliminated, the remaining non-Gal antigens are considered to be xenoantigens. Sialosyl-Tn or Hanganutziu-Deicher (HD), are non-Gal antigens specific to natural antibodies in human. To overcome rejection responses such as HAR, studies of genes involved in carbohydrate antigens, causing xenoantigenicity, are necessary. Knowledge of pig glycosyltransferases are also useful to apply to xenoantigen masking or identification of the xenoantigenic sialylglycan(s). In the first chapter the screening for pig glycosyltransferase genes for xenoantigens is presented. In the chapter II to IV the cloning, characterization, and investigation of the regulatory mechanism of the pig CMAH gene in NeuGc biosynthesis is shown. Lastly, the effects of an alteration of pig glycosylation patterns on human serum-mediated cytotoxicity, caused by human sialyltransferases including hST6GalNAc IV is presented.

1 Vascular Analysis of the Carotid Body in the Spontaneously Hypertensive Rat.- 2 Role of the Carotid Sinus Nerve and of Dopamine in the Biochemical Response of Sympathetic Tissues to Long-Term Hypoxia.- 3 The Effects of Almitrine on [3H]5HT and [125I] Endothelin Binding to Central and Peripheral Receptors: An In Vitro Autoradiographic Study in the Cat.- 4 Immunocytochemical and Neurochemical Aspects of Sympathetic Ganglion Chemosensitivity.- 5 Neuronal and Neuroendocrine Markers in the Human Carotid Body in Health and Disease.- 6 The Effects of Chronic Hypoxaemia upon the Structure of the Human Carotid Body.- 7 Dopaminergic and Peptidergic Sensory Innervation of the Rat Carotid Body: Organization and Development.- 8 Effects of Cell-Free Perfusion and Almitrine Bismesylate on the Ultrastructure of Type-1 Cell Mitochondria in the Cat Carotid Body.- 9 Multi-Unit Compartmentation of the Carotid Body Chemoreceptor by Perineurial Cell Sheaths: Immunohistochemistry and Freeze-Fracture Study.- 10 Light and Electronmicroscopical Immunohistochemical Investigation of the Innervation of the Human Carotid Body.- 11 Serotonin (5-Hydroxytryptamine) Expression in Pulmonary Neuro-Endocrine Cells (NE) and a Netumor Cell Line.- 12 Effects of Hypoxia on Cultured Chemoreceptors of the Rat Carotid Body: DNA Synthesis and Mitotic Activity in Glomus Cells.- 13 Localization of Dopamine D2 Receptor mRNA in the Rabbit Carotid Body and Petrosal Ganglion by in situ Hybridization.- 14 Noradrenergic Glomus Cells in the Carotid Body: An Autoradiographic and Immunocytochemical Study in the Rabbit and Rat.- 15 The Modulation of Intracellular pH in Carotid Body Glomus Cells by Extracellular pH and pCO2.- 16 Evidence for Glucose Uptake in the Rabbit Carotid Body.- 17 Effects of Inorganic Calcium Channel Blockers on Carotid Chemosensory Responses in the Cat.- 18 Those Strange Glomus Cells.- 19 Carotid Body Neurotransmission.- 20 Carbonic Anhydrase and the Carotid Body.- 21 Ca2+ Dynamics in Chemoreceptor Cells: An Overview.- 22 Spectrophotometric Analysis of Heme Proteins in Oxygen Sensing Cell Systems.- 23 Neurochemical and Molecular Biological Aspects on the Resetting of the Arterial Chemoreceptors in the Newborn Rat.- 24 Carbonic Anhydrase and Carotid Body Chemoreception in the Presence and Absence of CO2-HCO3-.- 25 Role of Ion-Exchangers in the Cat Carotid Body Chemotransduction.- 26 Dopamine Metabolism in the Rabbit Carotid Body in vitro: Effect of Hypoxia and Hypercapnia.- 27 PO2-Dependence of Phospholipase C in the Cat Carotid Body.- 28 Optical Measurements of Micro-Vascular Oxygen Pressure and Intracellular pH in the Cat Carotid Body: Testing Hypotheses of Oxygen Chemoreception.- 29 Elevation of Cytosolic Calcium Induced by pH Changes in Cultured Carotid Body Glomus Cells.- 30 Role of Carbon Dioxide for Hypoxic Chemotransduction of the Cat Carotid Body.- 31 Metabolic Substrate Dependence of Carotid Chemosensory Responses to Stop-Flow Evoked Hypoxia and to Nicotine.- 32 Effects of Chemosensory Stimulation Membrane Currents Recorded with the Perforated-Patch Method from Cultured Rat Glomus Cells.- 33 Carbonic Anhydrase Near Central Chemoreceptors.- 34 Update on the Bicarbonate Hypothesis.- 35 Regulation of Intracellular pH in Type I Cells of the Neonatal Rat Carotid Body.- 36 Noradrenergic Inhibition of the Goat Carotid Body.- 37 Role of Substance P in Rat Carotid Body Responses to Hypoxia and Capsaicin.- 38 Carotid Sinus Nerve Inhibition Mediated by Atrial Natriuretic Peptide.- 39 Neurotransmitters and Second Messenger Systems in the Carotid Body.- 40 Does Adenosine Stimulate Rat Carotid Body Chemoreceptors?.- 41 Effects of Haloperidol on Cat Carotid Body Chemoreceptionin Vitro.- 42 Effect of Arterial Chemoreceptor Stimulation: Role of Norepinephrine in Hypoxic Chemotransmission.- 43 Carotid Body Denervation and Pulmonary Vascular Resistance in the Rat.- 44 Effects of Chemoreceptor Stimulation by Almitrine Bismesylate on Renal Function in Conscious Rats...

This volume aims to provide an update on recent developments in protein secretion studies in plants versus yeast and mammalian systems. This book also discusses case studies that analyze the use of plant protein secretion using various tools and systems. The chapters in this book explore topics such as the study of Golgi-mediated protein traffic in plant cells; actin-based intracellular trafficking in pollen tubes; secretion system for identification of cargo proteins of vacuolar sorting receptors; isolation of the plant exocyst complex; and plant autophagy. 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. Comprehensive and authoritative, Plant Protein Secretion: Methods and Protocols is a valuable resource for researchers interested in furthering their studies in plant protein secretion."

Providing current diverse approaches and techniques used to study the immunoproteome, Immunoproteomics: Methods and Protocols collects chapters from key researchers that deliver information to be used in diagnostics, disease progression, and vaccine correlates of protection analysis, to name but a few. This detailed volume includes techniques used for the study of the antibody targets of bacterial pathogens, viruses, and cancer, mass spectrometry-based approaches to characterize T-cell epitopes, chapters on detection and relative quantification of cytokines in serum, as well as in silico prediction of epitopes using sequence-based or modeling approaches. 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. Practical and thorough, Immunoproteomics: Methods and Protocols aids researchers in transferring these techniques to their own laboratories in addition to providing a reference to guide researchers toward appropriate techniques.

Lectins are an important group of proteins, which bind to carbohydrates and can agglutinate cells or precipitate polysaccharides and glycoproteins. These agglutinating and precipitating properties are very similar to those of antibodies, and hence many of the methods used in lectin research are based on immunochemical techniques. Lectins are however not of immune origin, and are found in almost all organisms, including plants and micro-organisms. Current interest in lectins derives particularly from their usefulness in detecting and studying carbohydrates in solution and on cell surfaces. Studies of lectin-carbohydrate interactions are providing information on the precise molecular details of the interactions between proteins and carbohydrates in general. Lectins also serve as valuable tools in biological and medical research, in areas as diverse as separation and characterization of glycoproteins and glycopeptides, typing of bacteria and fractionation of lymphocytes and of bone marrow cells. Their biological role is uncertain but is generally believed to be primarily as recognition determinants in mirco-organisms, plants and animals. The purpose of the book is to give an accessible overview of the properties of lectins, their possible roles and their applications in biology and biomedical research in biochemistry and cell biology. This book should be of interest to biochemists, biologists, microbiologists, pathologists, histologists, cancer researchers, immunologists and haematologists.

Since its inception in 1945, this serial has provided critical and integrating articles written by research specialists that integrate industrial, analytical, and technological aspects of biochemistry, organic chemistry, and instrumentation methodology in the study of carbohydrates. The articles provide a definitive interpretation of the current status and future trends in carbohydrate chemistry and biochemistry.
Features contributions from leading authorities and industry experts Informs and updates on all the latest developments in the field

Carbohydrate Chemistry provides review coverage of all publications relevant to the chemistry of monosaccharides and oligosaccharides in a given year. The amount of research in this field appearing in the organic chemical literature is increasing because of the enhanced importance of the subject, especially in areas of medicinal chemistry and biology. In no part of the field is this more apparent than in the synthesis of oligosaccharides required by scientists working in glycobiology. Clycomedicinal chemistry and its reliance on carbohydrate synthesis is now very well established, for example, by the preparation of specific carbohydrate- based antigens, especially cancer-specific oligosaccharides and glycoconjugates. Coverage of topics such as nucleosides, amino-sugars, alditols and cyclitols also covers much research of relevance to biological and medicinal chemistry. Each volume of the series brings together references to all published work in given areas of the subject and serves as a comprehensive database for the active research chemist Specialist Periodical Reports provide systematic and detailed review coverage in major areas of chemical research. Compiled by teams of leading authorities in the relevant subject areas, the series creates a unique service for the active research chemist, with regular, in-depth accounts of progress in particular fields of chemistry. Subject coverage within different volumes of a given title is similar and publication is on an annual or biennial basis.

This book presents a common principle of actions of long noncoding RNAs (lncRNAs) from points of view at the atomic, molecular and cellular levels. At the atomic level, chemical studies of ribonucleic acids explain the chemical behavior of lncRNAs. Structural biological analysis of lncRNAs and its binding proteins also reveal the precise mechanisms of their actions. Molecular biological approaches lead to insights into molecular mechanisms of these lncRNA actions. At the cellular or individual level of analysis, we grasp the biology and medicine of lncRNAs. These three layers of approaches are thoroughly new and produce novel insights into functions of lncRNAs in living cells. The book consists of five parts: 1) Bioinformatics and other methodologies for lncRNAs, 2) Atomic and molecular structures of lncRNAs, 3) Molecular functions of lncRNAs, 4) Biological actions of lncRNAs, and 5) Potential outcomes for clinical medicine. These sections connect well and work synergistically. The book is for researchers whose specialty is RNA biology and chemistry and also for advanced students at the graduate and undergraduate levels. Readers can grasp the leading edge of lncRNA studies in a comprehensive manner and are inspired to pursue their own particular interests.

This new volume of "Methods in Enzymology" continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. Thethird of3 volumes covering Natural product biosynthesis by microorganisms and plants.
This new volume continues the legacy of this premier serialContains quality chapters authored by leaders in the fieldThe third of 3 volumes, it has chapters on such topics as metabolic pathways in "Aspergillus oryzae, "heterologous gene clusters and cyanobacteria as a source of natural products"

Cell biology spans among the widest diversity of methods in the biological sciences. From physical chemistry to microscopy, cells have given up with secrets only when the questions are asked in the right way This new volume of "Methods in Cell Biology" covers laboratory methods in cell biology, and includes methods that are among the most important and elucidating in the discipline, such as transfection, cell enrichment and magnetic batch separation.

Covers the most important laboratory methods in cell biology Chapters written by experts in their fields
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