Single-particle Cryo-electron Microscopy

Single-particle Cryo-electron Microscopy

Author: Joachim Frank

Publisher: World Scientific Publishing Company

Published: 2017-12-31

Total Pages: 0

ISBN-13: 9789813234857

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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.


Cryo-Electron Microscopy in Structural Biology

Cryo-Electron Microscopy in Structural Biology

Author: Krishnarao Appasani

Publisher: CRC Press

Published: 2024-10-17

Total Pages: 488

ISBN-13: 1040118844

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Cryo-electron microscopy, in combination with tomography, has emerged as a new technology for visualizing molecular structures at a resolution beyond even 1 Å. Using this technology has revealed the native molecular details of viruses, membranes, enzymes, ribosomes, and cells. This comprehensive volume brings together authoritative overviews of these methods from structural and biological perspectives. It is a must-have for researchers and graduate students, as well as those working in industry, primarily in the areas of biophysics, structural biology, crystallography, and genomics. Key Features • Focuses on the applications of cryo-EM to structural biology • Documents the importance of cryo-EM/ET approaches in studying the structural determinants of cellular organelle and membrane protein biochemistry • Reviews the applications of high-resolution structures of viruses • Emphasizes structural insights of nuclear and gene machineries • Includes a section focused entirely on the applications of cryo-EM/ET in drug discovery and therapeutic development


Correlative Imaging

Correlative Imaging

Author: Paul Verkade

Publisher: John Wiley & Sons

Published: 2019-11-04

Total Pages: 245

ISBN-13: 1119086450

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Brings a fresh point of view to the current state of correlative imaging and the future of the field This book provides contributions from international experts on correlative imaging, describing their vision of future developments in the field based on where it is today. Starting with a brief historical overview of how the field evolved, it presents the latest developments in microscopy that facilitate the correlative workflow. It also discusses the need for an ideal correlative probe, applications in proteomic and elemental analysis, interpretation methods, and how correlative imaging can incorporate force microscopy, soft x-ray tomography, and volume electron microscopy techniques. Work on placing individual molecules within cells is also featured. Correlative Imaging: Focusing on the Future offers in-depth chapters on: correlative imaging from an LM perspective; the importance of sample processing for correlative imaging; correlative light and volume EM; correlation with scanning probe microscopies; and integrated microscopy. It looks at: cryo-correlative microscopy; correlative cryo soft X-ray imaging; and array tomography. Hydrated-state correlative imaging in vacuo, correlating data from different imaging modalities, and big data in correlative imaging are also considered. Brings a fresh view to one of the hottest topics within the imaging community: the correlative imaging field Discusses current research and offers expert thoughts on the field’s future developments Presented by internationally-recognized editors and contributors with extensive experience in research and applications Of interest to scientists working in the fields of imaging, structural biology, cell biology, developmental biology, neurobiology, cancer biology, infection and immunity, biomaterials and biomedicine Part of the Wiley–Royal Microscopical Society series Correlative Imaging: Focusing on the Future will appeal to those working in the expanding field of the biosciences, correlative microscopy and related microscopic areas. It will also benefit graduate students working in microscopy, as well as anyone working in the microscopy imaging field in biomedical research.


Advances in Protein Molecular and Structural Biology Methods

Advances in Protein Molecular and Structural Biology Methods

Author: Timir Tripathi

Publisher: Academic Press

Published: 2022-01-14

Total Pages: 716

ISBN-13: 0323902650

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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


Biophysical Approaches for the Study of Membrane Structure Part A

Biophysical Approaches for the Study of Membrane Structure Part A

Author:

Publisher: Elsevier

Published: 2024-07-05

Total Pages: 536

ISBN-13: 0443293058

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Biophysical Approaches for the Study of Membrane Structure, Part A, Volume 700 explores lipid membrane asymmetry and lateral heterogeneity. A burst of recent research has shown that bilayers whose leaflets differ in their physical properties—such as composition, phase state, or lateral stress—exhibit many fascinating new characteristics, but also pose a host of new challenges related to their creation, characterization, simulation, and theoretical description. Chapters in this new release include Evaluation of functional transbilayer coupling in live cells by controlled lipid exchange and imaging FCS, Effects of lateral and hydrostatic pressure on membrane structure and properties, and much more.Other sections cover Using the yeast vacuole as a system to test the lipid drivers of membrane heterogeneity in living cells, Direct quantification of cellular membrane lipids using ratiometric fluorescence sensors, The spectral phasor approach to resolving membrane order with environmentally sensitive dyes, The use of hemifusion to create asymmetric giant unilamellar vesicles: Insights on induced order domains, Advanced microscopy methods to study membrane pores, Use of cryo-EM to study membrane phase separation, and much more. - Explore the state-of-the-art of lipid membrane asymmetry - Covers experimental, theoretical, and computational techniques to create and characterize asymmetric lipid membranes - Teaches how these kinds of approaches create and characterize laterally inhomogeneous membranes


Bacterial Secretion Systems

Bacterial Secretion Systems

Author: Laure Journet

Publisher: Springer Nature

Published: 2023-11-06

Total Pages: 607

ISBN-13: 1071634453

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This second edition details new and updated protocols that cover techniques used to study secretion systems. Chapters focus on identifying and localizing the different subunits, defining interactions within subunits, monitoring conformational changes, purifying and imaging of large complexes, defining the assembly pathway by fluorescence microscopy and the role of energy during assembly and/or secretion, identifying secreted effectors as well as using reporters to follow effector transport. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Bacterial Secretion Systems: Methods and Protocols- Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.


The Resolution Revolution: Recent Advances In cryoEM

The Resolution Revolution: Recent Advances In cryoEM

Author:

Publisher: Academic Press

Published: 2016-08-26

Total Pages: 488

ISBN-13: 0128054352

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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


Computational Drug Discovery

Computational Drug Discovery

Author: Pooja A. Chawla

Publisher: Walter de Gruyter GmbH & Co KG

Published: 2024-10-07

Total Pages: 543

ISBN-13: 3111207595

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Computational methods and understanding computational models are important in modern drug discovery. The book focuses on computational approaches that can improve the development of in silico methodologies. It includes lead hit methods, docking algorithms, computational chiral compounds, structure-based drug design, GROMACS and NAMD, structural genomics, toxicity prediction, enzyme inhibitors and peptidomimetic therapeutics


Structural Biology in Drug Discovery

Structural Biology in Drug Discovery

Author: Jean-Paul Renaud

Publisher: John Wiley & Sons

Published: 2020-01-09

Total Pages: 1437

ISBN-13: 1118900502

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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