This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. Methods to assess mitochondrial function is of great interest to neuroscientists studying chronic forms of neurodegeneration, including Parkinson's, Alzheimer's, ALS, Huntington's and other triplet repeat diseases, but also to those working on acute conditions such as stroke and traumatic brain injury. This volume covers research methods on how to assess the life cycle of mitochondria including trafficking, fusion, fission, and degradation. Multiple perspectives on the complex and difficult problem of measurement of mitochondrial reactive oxygen species production with fluorescent indicators and techniques ranging in scope from measurements on isolated mitochondria to non-invasive imaging of metabolic function.

This book provides a selection of recently developed methods and protocols in bacterial glycomics to aid in bettering our understanding of the structures and functions of bacterial polysaccharides, their attachments to proteins and lipids, their role in biofilm formation, as well as their biosynthesis. With the emerging bacterial resistance to commonly used antibiotics world-wide, these techniques to study the outer polysaccharides of bacteria, with their functions in bacterial adhesion, colonization, growth, establishment of biofilms, and control virulence and pathogenicity, are increasingly important. 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 cutting-edge, Bacterial Polysaccharides: Methods and Protocols aims to support researchers contributing to future approaches that will fill our knowledge gaps and define anti-bacterial targets.

This book provides a review of the multitude of nucleic acid polymerases, including DNA and RNA polymerases from Archea, Bacteria and Eukaryota, mitochondrial and viral polymerases, and other specialized polymerases such as telomerase, template-independent terminal nucleotidyl transferase and RNA self-replication ribozyme. Although many books cover several different types of polymerases, no book so far has attempted to catalog all nucleic acid polymerases. The goal of this book is to be the top reference work for postgraduate students, postdocs, and principle investigators who study polymerases of all varieties. In other words, this book is for polymerase fans by polymerase fans.

Nucleic acid polymerases play a fundamental role in genome replication, maintenance, gene expression and regulation. Throughout evolution these enzymes have been pivotal in transforming life towards RNA self-replicating systems as well as into more stable DNA genomes. These enzymes are generally extremely efficient and accurate in RNA transcription and DNA replication and share common kinetic and structural features. How catalysis can be so amazingly fast without loss of specificity is a question that has intrigued researchers for over 60 years. Certain specialized polymerases that play a critical role in cellular metabolism are used for diverse biotechnological applications and are therefore an essential tool for research.

First published in 1943, "Vitamins and Hormones" is the longest-running serial published by Academic Press.

The Series provides up-to-date information on vitamin and hormone research spanning data from molecular biology to the clinic. A volume can focus on a single molecule or on a disease that is related to vitamins or hormones. A hormone is interpreted broadly so that related substances, such as transmitters, cytokines, growth factors and others can be reviewed.

This volume focuses on endocrine disrupters.

Key features: Expertise of the contributorsCoverage of a vast array of subjectsIn depth current information at the molecular to the clinical levelsThree-dimensional structures in colorElaborate signaling pathways "

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 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"

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 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.

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.

This new volume of "Methods in Enzymology" continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. The second of3 volumes covering Natural product biosynthesis by microorganisms and plants.
This new volume continues the legacy of this premier serialContainsquality chapters authored by leaders in the field The second of3 volumes it has chapters on such topics as biological chlorination, bromination and iodination, and phylogenetic approaches to natural product structure prediction"

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"

This new volume of "Methods in Enzymology" continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. The volume covers nucleosomes, histones and chromatin and has chapters on dynamic mapping of histone-DNA interactions in nucleosomes by unzipping single molecules of DNA, digital DNase technology, and genome-wide analysis of chromatin transition.
Contains quality chapters authored by leaders in the fieldThe volume covers nucleosomes, histones and chromatinHas chapters on dynamic mapping of histone-DNA interactions in nucleosomes by unzipping single molecules of DNA, digital DNase technology, and genome-wide analysis of chromatin transition

This new volume of "Methods in Enzymology" continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. The first of 2 volumes covering nucleosomes, histones and chromatin, it has chapters on methods applied to the study of protein arginine methylation, high-resolution identification of intra- and interchromosomal DNA interactions by 4C technology, and peptide arrays to interrogate the binding specificity of chromatin-binding proteins.
Continues the legacy of this premier serial by containing quality chapters authored by leaders in the field The first of 2 volumes covering nucleosomes, histones and chromatin Chapters on methods applied to the study of protein arginine methylation, high-resolution identification of intra- and interchromosomal DNA interactions by 4C technology, and peptide arrays to interrogate the binding specificity of chromatin-binding proteins

Few problems in protein biochemistry have proven to be as challenging and recalcitrant as the molecular description of nitrogenase, the catalyst of one of the most remarkable chemical transformations in biological systems: the nucleotide-dependent reduction of atmospheric dinitrogen to bioavailable ammonia. In "Nitrogen Fixation: Methods and Protocols," recognized experts in the field provide an up-to-date, in-depth overview of the methods that have been applied to studying the nitrogenase at a molecular level, ranging from genetic, biochemical, spectroscopic, and chemical methods to theoretical calculations. In addition, techniques used to study an enzyme system that is homologous to nitrogenase are described in this book. Written in the highly successful "Methods in Molecular Biology " series format, methods chapters include introductions to their respective chapters, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Practical and cutting-edge, "Nitrogen Fixation: Methods and Protocols" will be useful for anyone interested in nitrogenase research and willing to venture further toward addressing the remaining mechanistic and biosynthetic questions of this fascinating enzyme system."

This volume of Methods in Enzymology is thesecond of 3 parts looking at current methodology for the imaging and spectroscopic analysis of live cells. The chapters provide hints and tricks not available in primary research publications. It is an invaluable resource for academics, researchers and students alike.
Expert authors who are leaders in the fieldExtensively referenced and useful figures and tablesProvides hints and tricks to facilitate reproduction of methods"

The combination of faster, more advanced computers and more quantitatively oriented biomedical researchers has recently yielded new and more precise methods for the analysis of biomedical data. These better analyses have enhanced the conclusions that can be drawn from biomedical data, and they have changed the way that experiments are designed and performed. This volume, along with the 2 previous "Computer Methods" volumes for the "Methods in Enzymology" serial, aims to inform biomedical researchers about recent applications of modern data analysis and simulation methods as applied to biomedical research.

* Presents step-by-step computer methods and discusses the techniques in detail to enable their implementation in solving a wide range of problems * Informs biomedical researchers of the modern data analysis methods that have developed alongside computer hardware *Presents methods at the "nuts and bolts" level to identify and resolve a problem and analyze what the results mean

State-of-the-art update on methods and protocols dealing with the detection, isolation and characterization of macromolecules and their hosting organisms that facilitate nitrification and related processes in the nitrogen cycle as well as the challenges of doing so in very diverse environments.

Provides state-of-the-art update on methods and protocols Deals with the detection, isolation and characterization of macromolecules and their hosting organisms deals with the challenges of very diverse environments.

Proteases occur naturally in all organisms. They are enzymes that are involved in many physiological reactions such as digestion of food and blood clotting. This volume reviews their role in health and disease and presents the latest research and developments.

* Discusses new discoveries, approaches, and ideas * Contributions from leading scholars and industry experts * Reference guide for researchers involved in molecular biology and related fields

This volume of "The Enzymes" features high-caliber thematic articles on the topic of glycosylphosphatidylinositol (GPI) anchoring of proteins.
* Contributions from leading authorities
* Informs and updates on all the latest developments in the field

Multicellular organisms must be able to adapt to cellular events to accommodate prevailing conditions. Sensory-response circuits operate by making use of a phosphorylation control mechanism known as the "two-component system." This volume, the third in a three-volume treatment edited by the same group of editors, includes a wide range of methods, including those dealing with the Sln-1 kinase pathway, triazole sensitivity in "C. albicans," and histidine kinases in cyanobacteria circadian clock.

* Includes time-tested core methods and new innovations applicable to any researcher studing two-component signaling systemsor histidine kinases * Methods included are useful to both established researchers and newcomers to the field * Relevant background and reference information given for procedures can be used as a guide to developing protocols in a number of disciplines"

This volume of "The Enzymes" features high-caliber thematic articles on the topic of glycosylphosphatidylinositol (GPI) anchoring of proteins.
* Contributions from leading authorities
* Informs and updates on all the latest developments in the field

Kinetic studies of enzyme action provide powerful insights into the underlying mechanisms of catalysis and regulation. These approaches are equally useful in examining the action of newly discovered enzymes and therapeutic agents.
Contemporary Enzyme Kinetics and Mechanism, Second Edition presents key articles from Volumes 63, 64, 87, 249, 308 and 354of Methods in Enzymology. The chapters describe the most essential and widely applied strategies. A set of exercises and problems is included to facilitate mastery of these topics.
The book will aid the reader to design, execute, and analyze kinetic experiments on enzymes. Its emphasis on enzyme inhibition will also make it attractive to pharmacologists and pharmaceutical chemists interested in rational drug design.
Of the seventeen chapters presented in this new edition, ten did not previously appear in the first edition.
Key Features
* Transient kinetic approaches to enzyme mechanisms
* Designing initial rate enzyme assay
* Deriving initial velocity and isotope exchange rate equations
* Plotting and statistical methods for analyzing rate data
* Cooperativity in enzyme function
* Reversible enzyme inhibitors as mechanistic probes
* Transition-state and multisubstrate inhibitors
* Affinity labeling to probe enzyme structure and function
* Mechanism-based enzyme inactivators
* Isotope exchange methods for elucidating enzymatic catalysis
* Kinetic isotope effects in enzyme catalysis
* Site-directed mutagenesis in studies of enzyme catalysis"

Microbial natural products have been an important traditional source of valuable antibiotics and other drugs but interest in them waned in the 1990s when big pharma decided that their discovery was no longer cost-effective and concentrated instead on synthetic chemistry as a source of novel compounds, often with disappointing results. Moreover understanding the biosynthesis of complex natural products was frustratingly difficult. With the development of molecular genetic methods to isolate and manipulate the complex microbial enzymes that make natural products, unexpected chemistry has been revealed and interest in the compounds has again flowered. This two-volume treatment of the subject will showcase the most important chemical classes of complex natural products: the peptides, made by the assembly of short chains of amino acid subunits, and the polyketides, assembled from the joining of small carboxylic acids such as acetate and malonate. In both classes, variation in sub-unit structure, number and chemical modification leads to an almost infinite variety of final structures, accounting for the huge importance of the compounds in nature and medicine.
* Gathers tried and tested methods and techniques from top players in the field.
* In depth coverage of ribosomally-synthesised and Non-ribosomally-synthesised peptides.
* Provides an extremely useful reference for the experienced research scientist.

Ribonucleases are a ubiquitous and functionally diverse group of enzymes that have a common ability to cleave RNA. Either through scission of internal phosphodiesters, or removal of nucleotides from RNA 5' or 3' ends, ribonucleases perform essential roles in gene expression and regulation, genome replication and maintenance, host defense, stress response, and viral strategies of infection. Ribonucleases have also served as highly informative models to understand virtually every aspect of biomolecular structure and function. The fifteen chapters in this volume are written by recognized researchers in the field, and provide in-depth analyses of the major ribonuclease families. Particular focus is given to the relation of ribonuclease structure and mechanism to biological function, as well as ribonuclease dysfunction in certain disease states. Other topics include the evolutionary genetics and functional diversification of ribonucleases, engineered ribonucleases as anti-cancer agents, the mechanisms of action of artificial ribonucleases, and ribonucleases as models to understand protein folding and stability. This volume should serve as an essential reference for a broad range of researchers and educators with interests in RNA metabolism, enzymology, and gene regulation.