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This updated and expanded volume reflects the current state of the structural protein field with improved and refined protocols that have been applied to particularly challenging proteins. Beginning with a section on structural bioinformatics, the book continues with sections covering the challenge of producing high quality samples for structural studies, particularly mammalian membrane proteins and protein complexes, as well as protocols for structure determination, including the use of electrons in structural biology and more. 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 up-to-date, Structural Proteomics: High-Throughput Methods, Third Edition will aid researchers in expanding our knowledge of this vital and expansive area of protein science.
Chapter 9 is available open access under a CC BY 4.0 license.
List of contents
The Protein Data Bank Archive.- Computational Methods for the Elucidation of Protein Structure and Interactions.- Methods for Molecular Modeling of Protein Complexes.- High-Level Production of Recombinant Eukaryotic Proteins from Mammalian Cells Using Lentivirus.- Transient Transfection and Expression of Eukaryotic Membrane Proteins in Expi293F Cells and Their Screening on a Small-Scale: Application for Structural Studies.- Reproducible and Easy Production of Mammalian Proteins by Transient Gene Expression in High Five Insect Cells.- SynBac: Enhanced Baculovirus Genomes by Iterative Recombineering.- Gene Tagging with the CRISPR-Cas9 System to Facilitate Macromolecular Complex Purification.- Synthesis of Fluorescently Labeled Antibodies Using Non-Canonical Amino Acids in Eukaryotic Cell-Free Systems.- Solid-State NMR Spectroscopy for Studying Microtubules and Microtubule-Associated Proteins.- Dynamic Structural Biology Experiments at XFEL or Synchrotron Sources.- From Tube to Structure:SPA Cryo-EM Workflow Using Apoferritin as an Example.- Image Processing in Cryo-Electron Microscopy of Single Particles: The Power of Combining Methods.- Setup and Troubleshooting of Volta Phase Plate Cryo-EM Data Collection.- Cryo-Focused Ion Beam Lamella Preparation Protocol for In Situ Structural Biology.- Protein and Small Molecule Structure Determination by the Cryo-EM Method MicroED.
