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
Editor
List of contributors
Foreword
Preface
SECTION I Cryo-EM in Cell Biology: Structure Determination of Protein
Assemblies and Cellular Organelle
Chapter 1 Introduction on Cryogenic Electron Microscopy in Structural
Biology
Krishnarao Appasani
Chapter 2 Structural Dynamics of Proteasome Regulation by Cryo-EM
Youdong Mao
Chapter 3 Cryo-EM Structures of a Ubiquitin-Specific Protease in Complex with
Substrate
Martin Rennie
Chapter 4 Structural Biology of Protein Kinases and Their Regulation
Samrat Mitra, Pranita Ray, Prateeka Borar, and Smarajit Polley
Chapter 5 The Life and Times of p53: Structural Advancements to Decipher the
Guardian of the Genome
Maria J. Solares, Samantha Berry, and Deborah F. Kelly
SECTION II Technology Platform and Evolving Technologies
Chapter 6 Single-Particle Cryo-EM: What Happens Inside the Black Box?
Sayan Bhakta and Marin van Heel
Chapter 7 Streptavidin Affinity Grids in the Study of Macromolecular
Complexes Using Cryo-Electron Microscopy
Nils Marechal and Patrick Schultz
Chapter 8 Running a Cost-Effective Modern Cryo-EM Facility for SPA Analysis
Lijia Jia
Chapter 9 Computational Methods for Bimolecular Structure Modeling for
Cryo-EM
Javad Baghirov, Xiao Wang, Genki Terashi, Shu Li, and Daisuke Kihara
Chapter 10 Artificial Intelligence-Driven Data Acquisition for
Single-Particle Cryo-EM and Three-Dimensional Electron Diffraction
Koji Yonekura
Chapter 11 From Icy Specimens to Digital Insights: Integrating Sample
Preparation, Data Analysis, and IT Infrastructure
Eric Shell, Karine Minari, and Vitor Hugo Balasco Serrćo
SECTION III Cryo-EM in Virology and Enzymology: High-Resolution Structures of
Chaperonins and Enzymes
Chapter 12 Cryo-EM Structures of Viral and Bacterial Chaperonins
Olga S. Sokolova and Tatiana B. Stanishneva-Konovalova
Chapter 13 Cryo-EM Structure of Monkeypox Virus DNA Polymerase Holoenzyme
Yi Shi and George F Gao
Chapter 14 Determining the High-Resolution Structure of the
[ NiFe]-hydrogenase Huc by Native Host Purification and Cryo-EM
Hari Venugopal, Ashleigh Kropp, Chris Greening, and Rhys Grinter
Chapter 15 Membrane Rupture and LipidProtein Interaction of -Hemolysin from
Staphylococcus aureus Using Cryo-EM
Suman Mishra, Anupam Roy, Arnab Chatterjee, and Somnath Dutta
SECTION IV Cryo-EM in the Study of Membrane Biology: Probing the Structures
of Membrane Proteins
Chapter 16 Historical Evolution and Perspective on Cryo-EM Analysis of
Membrane Proteins
Youzhong Guo
Chapter 17 Single-Particle Cryogenic Electron Microscopy Unfolds Distinct
Membrane Transporter
Folds: Focus on Autoinducer-2 Exporter Family
Radhika Khera
Chapter 18 Rhodopsin Dimers in the Retinal Rod Outer Segment
Sahil Gulati and Krzysztof Palczewski
Chapter 19 Cryo-EM Structures of the Fungal GPCR Dimer Ste2
Vaithish Velazhahan and Christopher G. Tate
Chapter 20 Human V-ATPases: Assembly, Function, and Regulation
Elizabeth Fosuah, Qingpeng Lin, and Tian-Min Fu
Chapter 21 Structure and Function of the Intracellular Calcium Channel
Ryanodine Receptor
Marco C. Miotto
Chapter 22 Cryo-EM of Mitochondrial Membrane Proteins
Swati Yadav, Shaileshanand Jha, and Kutti R. Vinothkumar
SECTION V Cryo-EM in the Study of Molecular Biology: Structural Insights of
Gene Machineries
Chapter 23 Imaging Structurally Dynamic Ribosomes with Cryogenic Electron
Microscopy
Samantha M. Webster, Mira B. May, Barrett M. Powell, and Joseph H. Davis
Chapter 24 Cryo-EM in the Study of Ribosome Biogenesis
Ankita Arora and Prem S. Kaushal
Chapter 25 Cryo-EM Structures of Ribosomes from Human Pathogens: Implications
for Drug Targeting
Ankit Dhur, Aneek Banerjee, Krishnamoorthi Srinivasan, and Jayati Sengupta
SECTION VI Cryo-EM in the Study of Neurobiology and SkinNeuroreceptors to
Ion Channels and Synaptic Transmission
Chapter 26 Cryo-EM Structural Features of Glutamate Receptors Involved in
Synaptic Neurotransmission or Brain Pathology
Neboja Bogdanovi
Chapter 27 Cryo-EM Structures of Systemic Amyloid Fibrils
Binh An Nguyen, Maria del Carmen Fernandez-Ramirez, Shumaila Afrin, and
Lorena Saelices
Chapter 28 Structural Insights into the 5-HT3A Receptor Revealed by Cryogenic
Electron Microscopy (Cryo-EM)
Zhuowen Li, Nikhil Bharambe, and Sandip Basak
Chapter 29 Molecular Cryo-EM in the Study of Skin Tissue
Lars Norlén
SECTION VII Applications of Cryo-ET and Cryo-EM in the Biopharmaceutical
Industry
Chapter 30 Emerging Technologies in Cryo-Electron Tomography
Kanika Khanna
Chapter 31 Cryo-Electron Tomographic Imaging of Membrane Proteins from
Mitochondria and Chloroplasts
Tofayel Ahmed and Antara Saha
Chapter 32 Functional Studies of the Actin Cytoskeleton by Cryogenic Electron
Tomography
Dorit Hanein and Niels Volkmann
Chapter 33 Cryo-EM in Drug Discovery
Youxin Kong, Maxime Killer, Madhumati Sevvana, and Chiara Rapisarda
Chapter 34 Applications of Single-Particle Analysis and Cryo-ET in Drug
Discovery
Qing Yao
Index
Krishnarao Appasani trained as a molecular cell biologist, earning Masters and Doctoral degrees in Biochemistry and Molecular biology. He accepted a Lectureship at Boston University and taught cell and molecular biology courses to undergraduates and graduate students. He was a post-doc in the laboratory of Nobel Laureate, the late Prof. H. Gobind Khorana at the Massachusetts Institute of Technology. Subsequently, he joined the faculty of Harvard Medical School and directed the Thoracic Oncology Laboratory at the Brigham &Womens Hospital and the Dana-Farber Cancer Institute in Boston. During this time he completed an MBA at Bryant University, Smithfield, RI, and then moved to Perkin Elmer and directed the Microarrays group. Dr Appasani has published about 60 research papers and abstracts in international journals, especially on Gene Expression. He was awarded the Outstanding Investigator Award by the American Federation for Medical Research, Washington D.C., competed globally for the Ranbaxy Research Medal, and was honored with the 1997-Ranbaxy Research Award in the field of Applied Medical Sciences. At Perkin Elmer, Dr Appasani organized a global Biomics Seminar series to educate scientists and to initiate and develop collaborations with researchers in academia and other industries. For about 15 years he ran a multi-million-dollar knowledge management enterprise and served as an international authority in bridging Academia and Industry by fostering inter-disciplinary and translational science. In 2016 Dr Appasani was inducted as a Life Fellow of the Royal Society of Biology, England, for his global efforts in promoting science education.
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