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This edited book is written for students, postdocs and established investigators who want to enter the field of single-particle cryo-EM. This is a recently developed method to determine high-resolution structures of biological macromolecules. A major strength is the fact that cryo-EM does not require prior crystallization of protein complexes. It is especially well suited for larger complexes and molecular machines. This book, provides a comprehensive, accessible and authoritative introduction to the field. It covers all necessary background, ranging from the underlying concepts to practical aspects such as specimen preparation, data-collection, data analysis, and the final validation of results. Key features Written for students, postdocs and established investigators who want to enter the field of single-particle cryo-EM Provides a comprehensive, accessible and authoritative introduction to the field of high-resolution structure analysis by single-article cryo-EM Covers all necessary background, ranging from the underlying concepts to practical aspects such as specimen preparation, data-collection, data analysis, and the final validation of results Authors of individual sections of this book have been recruited from among the most authoritative leaders in each topic
The book reproduces 55 of more than 300 articles written by the author, representing milestones in methods development of single-particle cryo-EM as well as important results obtained by this technique in the study of biological macromolecules and their interactions. Importantly, neither symmetries nor ordered arrangements (as in two-dimensional crystals, helical assemblies, icosahedral viruses) are required. Although the biological applications are mainly in the area of ribosome structure and function, the elucidation of membrane channel structures and their activation and gating mechanisms are represented, as well. The book is introduced by a commentary that explains the original development of concepts, describes the contributions of the author's colleagues and students, and shows how challenges were overcome as the technique matured. Along the way, the ribosome served as an example for a macromolecule with intricate structure and conformational dynamics that pose challenges for three-dimensional visualization. Toward the end of the book -- bringing us to the present time -- molecular structures with near-atomic resolution are presented, and a novel type of computational analysis, manifold embedding, is introduced. Single-particle cryo-EM is currently revolutionizing structural biology, presenting a powerful alternative to X-ray crystallography as a means to solve the structure of biological macromolecules. The book presents in one place a number of articles containing key advances in mathematical and computational methods leading up to the present time. Secondly, the development of the technique over the years is reflected by ever-expanding discoveries in the field of ribosome structure and function. Thirdly, as all histories of ideas, the history of concepts pertaining to this new method of visualization is fascinating all in itself.
Biological Macromolecules: Bioactivity and Biomedical Applications presents a comprehensive study of biomacromolecules and their potential use in various biomedical applications. Consisting of four sections, the book begins with an overview of the key sources, properties and functions of biomacromolecules, covering the foundational knowledge required for study on the topic. It then progresses to a discussion of the various bioactive components of biomacromolecules. Individual chapters explore a range of potential bioactivities, considering the use of biomacromolecules as nutraceuticals, antioxidants, antimicrobials, anticancer agents, and antidiabetics, among others. The third section of the book focuses on specific applications of biomacromolecules, ranging from drug delivery and wound management to tissue engineering and enzyme immobilization. This focus on the various practical uses of biological macromolecules provide an interdisciplinary assessment of their function in practice. The final section explores the key challenges and future perspectives on biological macromolecules in biomedicine. - Covers a variety of different biomacromolecules, including carbohydrates, lipids, proteins, and nucleic acids in plants, fungi, animals, and microbiological resources - Discusses a range of applicable areas where biomacromolecules play a significant role, such as drug delivery, wound management, and regenerative medicine - Includes a detailed overview of biomacromolecule bioactivity and properties - Features chapters on research challenges, evolving applications, and future perspectives
Three-Dimensional Electron Microscopy of Macromolecular Assemblies is the first systematic introduction to single-particle methods of reconstruction. It covers correlation alignment, classification, 3D reconstruction, restoration, and interpretation of the resulting 3D images in macromolecular assemblies. It will be an indispensable resource for newcomers to the field and for all using or adopting these methods.Key Features* Presents methods that offer an alternative to crystallographic techniques for molecules that cannot be crystallized* Describes methods that have been instrumental in exploring the three-dimensional structure of* the nuclear pore complex* the calcium release channel;* the ribosome* chaperonins
With the most comprehensive and up-to-date overview of structure-based drug discovery covering both experimental and computational approaches, Structural Biology in Drug Discovery: Methods, Techniques, and Practices describes principles, methods, applications, and emerging paradigms of structural biology as a tool for more efficient drug development. Coverage includes successful examples, academic and industry insights, novel concepts, and advances in a rapidly evolving field. The combined chapters, by authors writing from the frontlines of structural biology and drug discovery, give readers a valuable reference and resource that: Presents the benefits, limitations, and potentiality of major techniques in the field such as X-ray crystallography, NMR, neutron crystallography, cryo-EM, mass spectrometry and other biophysical techniques, and computational structural biology Includes detailed chapters on druggability, allostery, complementary use of thermodynamic and kinetic information, and powerful approaches such as structural chemogenomics and fragment-based drug design Emphasizes the need for the in-depth biophysical characterization of protein targets as well as of therapeutic proteins, and for a thorough quality assessment of experimental structures Illustrates advances in the field of established therapeutic targets like kinases, serine proteinases, GPCRs, and epigenetic proteins, and of more challenging ones like protein-protein interactions and intrinsically disordered proteins
cryoEM, a new volume in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods and new developments in recording images, the creation, evaluation and validation of 3D maps from the images, model building into maps and refinement of the resulting atomic structures, and applications of essentially single particle methods to helical structures and to sub-tomogram averaging. - Continues the legacy of this premier serial with quality chapters authored by leaders in the field - Covers research methods that determine the structures of biological molecules, a vital step for understanding their function - Contains the technical developments underpinning the advances of cryoEM and captures the exciting insights that have resulted
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
Electron microscopy has revolutionized our understanding the extraordinary intellectual demands required of the mi of materials by completing the processing-structure-prop croscopist in order to do the job properly: crystallography, erties links down to atomistic levels. It now is even possible diffraction, image contrast, inelastic scattering events, and to tailor the microstructure (and meso structure ) of materials spectroscopy. Remember, these used to be fields in them to achieve specific sets of properties; the extraordinary abili selves. Today, one has to understand the fundamentals ties of modem transmission electron microscopy-TEM of all of these areas before one can hope to tackle signifi instruments to provide almost all of the structural, phase, cant problems in materials science. TEM is a technique of and crystallographic data allow us to accomplish this feat. characterizing materials down to the atomic limits. It must Therefore, it is obvious that any curriculum in modem mate be used with care and attention, in many cases involving rials education must include suitable courses in electron mi teams of experts from different venues. The fundamentals croscopy. It is also essential that suitable texts be available are, of course, based in physics, so aspiring materials sci for the preparation of the students and researchers who must entists would be well advised to have prior exposure to, for carry out electron microscopy properly and quantitatively.
The present volume continues the trend established in previous volumes in this series on Advances in Structural Biology. As in the past, diverse topics of current importance relevant to the theme of the series are included in the fourth volume.
This volume details the most up-to-date cryo-EM techniques from leading researchers. Chapters are organized into four parts with emphasis on electron cryotomography, single particle analysis, and the crystal based cryo-EM methods of 2D electron crystallography, and MicroED for the study of 3D crystals. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, CryoEM: Methods and Protocols aims to serve as an excellent resource on cryo-EM and can serve as the foundation for new researchers to this growing field in structural biology.