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Scattering Methods in Structural Biology, Part B, Volume 676 in the Methods in Enzymology serial, highlights advances in the field, presenting chapters on Quality controls, Refining biomolecular structures and ensembles by SAXS-driven molecular dynamics simulations, Data analysis and modelling of small-angle scattering data with contrast variation, Observing protein degradation in solution by the PAN-20S proteasome complex: state-of-the-art and future perspectives of TR-SANS as a complementary tool to NMR, crystallography and Cryo-EM, Extracting structural insights from chemically-specific soft X-ray scattering, Reconstruction of 3D density of biological macromolecules from solution scattering, ATSAS- present state and new developments in computational methods, and much more. Additional chapters cover Modeling Structure and Dynamics of Protein Complexes with SAXS Profiles (FoXSDock and MultiFoXS), Validation of macromolecular flexibility in solution by SAXS, Combining NMR, SAXS and SANS to characterize the structure and dynamics of protein complexes, Application of Molecular Simulation Methods to Analyze SAS Data, and more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Enzymology serial - Updated release includes the latest information on Small Angle Scattering Methods for Structural Interpretation
This book provides a clear, comprehensible and up-to-date description of how Small Angle Scattering (SAS) can help structural biology researchers. SAS is an efficient technique that offers structural information on how biological macromolecules behave in solution. SAS provides distinct and complementary data for integrative structural biology approaches in combination with other widely used probes, such as X-ray crystallography, Nuclear magnetic resonance, Mass spectrometry and Cryo-electron Microscopy. The development of brilliant synchrotron small-angle X-ray scattering (SAXS) beam lines has increased the number of researchers interested in solution scattering. SAS is especially useful for studying conformational changes in proteins, highly flexible proteins, and intrinsically disordered proteins. Small-angle neutron scattering (SANS) with neutron contrast variation is ideally suited for studying multi-component assemblies as well as membrane proteins that are stabilized in surfactant micelles or vesicles. SAS is also used for studying dynamic processes of protein fibrillation in amyloid diseases, and pharmaceutical drug delivery. The combination with size-exclusion chromatography further increases the range of SAS applications. The book is written by leading experts in solution SAS methodologies. The principles and theoretical background of various SAS techniques are included, along with practical aspects that range from sample preparation to data presentation for publication. Topics covered include techniques for improving data quality and analysis, as well as different scientific applications of SAS. With abundant illustrations and practical tips, we hope the clear explanations of the principles and the reviews on the latest progresses will serve as a guide through all aspects of biological solution SAS. The scope of this book is particularly relevant for structural biology researchers who are new to SAS. Advanced users of the technique will find it helpful for exploring the diversity of solution SAS methods and applications. Chapter 3 of this book is available open access under a CC BY 4.0 license at link.springer.com.
Small Angle Scattering, Part A: Methods for Structural Investigation, Volume 675 in the Methods in Enzymology series, highlights new advances in the field, with new chapters in this updated release including SAXS foundations and metrics, Contrast variation sample preparation protocols, experimental procedures, and rudimentary analysis, Molecular deuteration for neutron scattering, Planning, Executing and Assessing the Feasibility of SANS Contrast Variation Experiments, Technical considerations for small-angle neutron scattering from biological macromolecules, and Advanced sample environments and capabilities at our synchrotron X-ray beamline with example applications. Additional sections in the book cover SEC-SAXS-MALS data acquisition and processing pipeline at SIBYLS, SEC-SAXS: pros and cons, experimental set-up, examples and software developments, Radiation damage and sample economy for stopped-flow methods in the time regime of millisecond and above, Stopped-flow-time-resolved SAXS, Insights on Temp-jump, time-resolved SAXS, and much more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Enzymology series - Includes the latest information on Small Angle Scattering: Methods for Structural Investigation
Biochemical Pathways and Environmental Responses in Plants, Part B, Volume 682 in the Methods in Enzymology series, highlights advances in the field with this new volume presenting chapters on MIE 681/682: Biochemical pathways and environmental responses in plants, Structure, function, and engineering of plant polyketide synthases, A sensitive LC-MS/MS assay for enzymatic characterization of methylthioalkylmalate synthase involved in glucosinolate side-chain elongation, Assaying formate-tetrahydrofolate ligase with monoglutamylated and polyglutamylated substrates using a fluorescence-HPLC based assay, An Approach to Nearest Neighbor Analysis of Pigmented Protein Complexes by Using Chemical Crosslinking in Combination with Mass Spectrometry, Biochemical characterization of plant aromatic aminotransferases, and much more. Other chapters focus on Functional Analysis of Phosphoethanolamine N-methyltransferase (PMT) in Plants and Parasites, A structure-guided computational screening approach for predicting plant enzyme-metabolite interactions, Plant metacaspase: an example of microcrystal structure determination and analysis, Biocatalytic system for comparative assessment of functional association of cytochrome P450 monooxygenases with their redox partners, Dirigent Protein Family Function and Structure, and more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in Methods in Enzymology series - Includes the latest information on Biochemical pathways and environmental responses in plants
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." Sections in Two-Component Signaling Systems, Part B include: - Structural Approaches - Reconstitution of Heterogeneous Systems - Intracellular Methods and Assays - Genome-Wide Analyses of Two-Component Systems - Presents detailed protocols - Includes troubleshooting tips
Advances in Protein Molecular and Structural Biology Methods offers a complete overview of the latest tools and methods applicable to the study of proteins at the molecular and structural level. The book begins with sections exploring tools to optimize recombinant protein expression and biophysical techniques such as fluorescence spectroscopy, NMR, mass spectrometry, cryo-electron microscopy, and X-ray crystallography. It then moves towards computational approaches, considering structural bioinformatics, molecular dynamics simulations, and deep machine learning technologies. The book also covers methods applied to intrinsically disordered proteins (IDPs)followed by chapters on protein interaction networks, protein function, and protein design and engineering. It provides researchers with an extensive toolkit of methods and techniques to draw from when conducting their own experimental work, taking them from foundational concepts to practical application. - Presents a thorough overview of the latest and emerging methods and technologies for protein study - Explores biophysical techniques, including nuclear magnetic resonance, X-ray crystallography, and cryo-electron microscopy - Includes computational and machine learning methods - Features a section dedicated to tools and techniques specific to studying intrinsically disordered proteins
SMALL-ANGLE SCATTERING A comprehensive and timely volume covering contemporary research, practical techniques, and theoretical approaches to SAXS and SANS Small-Angle Scattering: Theory, Instrumentation, Data, and Applications provides authoritative coverage of both small-angle X-ray scattering (SAXS), small-angle neutron scattering (SANS) and grazing incidence small-angle scattering (GISAS) including GISAXS and GISANS. This single-volume resource offers readers an up-to-date view of the state of the field, including the theoretical foundations, experimental methods, and practical applications of small-angle scattering (SAS) techniques including laboratory and synchrotron SAXS and reactor/spallation SANS. Organized into six chapters, the text first describes basic theory, instrumentation, and data analysis. The following chapters contain in-depth discussion on various applications of SAXS and SANS and GISAXS and GISANS, and on specific techniques for investigating structure and order in soft materials, biomolecules, and inorganic and magnetic materials. Author Ian Hamley draws from his more than thirty years’ experience working with many systems, instruments, and types of small-angle scattering experiments across most European facilities to present the most complete introduction to the field available. This book: Presents uniquely broad coverage of practical and theoretical approaches to SAXS and SANS Includes practical information on instrumentation and data analysis Offers useful examples and an accessible and concise presentation of topics Covers new developments in the techniques of SAXS and SANS, including GISAXS and GISANS Small-Angle Scattering: Theory, Instrumentation, Data, and Applications is a valuable source of detailed information for researchers and postgraduate students in the field, as well as other researchers using X-ray and neutron scattering to investigate soft materials, other nanostructured materials and biomolecules such as proteins.
This MIE volume provides laboratory techniques that aim to predict the structure of a protein which can have tremendous implications ranging from drug design, to cellular pathways and their dynamics, to viral entry into cells. - Expert researchers introduce the most advanced technologies and techniques in protein structure and folding - Includes techniques on tiling assays
The definitive guide to mass spectrometry techniques in biology and biophysics The use of mass spectrometry (MS) to study the architecture and dynamics of proteins is increasingly common within the biophysical community, and Mass Spectrometry in Structural Biology and Biophysics: Architecture, Dynamics, and Interaction of Biomolecules, Second Edition provides readers with detailed, systematic coverage of the current state of the art. Offering an unrivalled overview of modern MS-based armamentarium that can be used to solve the most challenging problems in biophysics, structural biology, and biopharmaceuticals, the book is a practical guide to understanding the role of MS techniques in biophysical research. Designed to meet the needs of both academic and industrial researchers, it makes mass spectrometry accessible to professionals in a range of fields, including biopharmaceuticals. This new edition has been significantly expanded and updated to include the most recent experimental methodologies and techniques, MS applications in biophysics and structural biology, methods for studying higher order structure and dynamics of proteins, an examination of other biopolymers and synthetic polymers, such as nucleic acids and oligosaccharides, and much more. Featuring high-quality illustrations that illuminate the concepts described in the text, as well as extensive references that enable the reader to pursue further study, Mass Spectrometry in Structural Biology and Biophysics is an indispensable resource for researchers and graduate students working in biophysics, structural biology, protein chemistry, and related fields.
Current techniques for studying biological macromolecules and their interactions are based on the application of physical methods, ranging from classical thermodynamics to more recently developed techniques for the detection and manipulation of single molecules. Reflecting the advances made in biophysics research over the past decade, and now including a new section on medical imaging, this new edition describes the physical methods used in modern biology. All key techniques are covered, including mass spectrometry, hydrodynamics, microscopy and imaging, diffraction and spectroscopy, electron microscopy, molecular dynamics simulations and nuclear magnetic resonance. Each method is explained in detail using examples of real-world applications. Short asides are provided throughout to ensure that explanations are accessible to life scientists, physicists and those with medical backgrounds. The book remains an unparalleled and comprehensive resource for graduate students of biophysics and medical physics in science and medical schools, as well as for research scientists looking for an introduction to techniques from across this interdisciplinary field.