Sameer Velankar, Stephen K. Burley, Genji Kurisu, Jeffery C. Hoch, and John L. Markley
2. Computational Methods for the Elucidation of Protein Structure and Interactions
Nicholas S. Edmunds and Liam J. McGuffin
3. Methods for Molecular Modeling of Protein Complexes
Tejashree Rajaram Kanitkar,, Neeladri Sen, Sanjana Nair, Neelesh Soni, Kaustubh Amritkar, Yogendra Ramtirtha, and M.S. Madhusudhan
Part II: Protein Production
4. High-Level Production of Recombinant Eukaryotic Proteins from Mammalian Cells Using Lentivirus
Ester Behiels and Jonathan Elegheert
5. Transient Transfection and Expression of Eukaryotic Membrane Proteins in Expi293F Cells and Their Screening on a Small-Scale: Application for Structural Studies
Ganna O. Krasnoselska, Maud Dumoux, Nadisha Gamage, Harish Cheruvara, James Birch, Andrew Quigley, and Raymond J. Owens
6. Reproducible and Easy Production of Mammalian Proteins by Transient Gene Expression in High Five Insect Cells
Maren Schubert, Manfred Nimtz, Federico Bertoglio, Stefan Schmelz, Peer Lukat, and Joop van den Heuvel
7. SynBac: Enhanced Baculovirus Genomes by Iterative Recombineering
Hannah Crocker, Barbara Gorda, Martin Pelosse, Deepak Balaji Thimiri Govinda Raj, and Imre Berger
8. Gene Tagging with the CRISPR-Cas9 System to Facilitate Macromolecular Complex Purification
Sylvain Geny, Simon Pichard, Arnaud Poterszman, and Jean-Paul Concordet
9. Synthesis of Fluorescently Labeled Antibodies Using Non-Canonical Amino Acids in Eukaryotic Cell-Free Systems
Marlitt Stech, Nathanaël Rakotoarinoro, Tamara Teichmann, Anne Zemella, Lena Thoring, and Stefan Kubick
Part III: Structure Determination
10. Solid-State NMR Spectroscopy for Studying Microtubules and Microtubule-Associated Proteins
Yanzhang Luo, Shengqi Xiang, Alessandra Lucini Paioni, Agnes Adler, Peter Jan Hooikaas, Jijumon A.S., Carsten Janke3, Anna Akhmanova, and Marc Baldus
11. Dynamic Structural Biology Experiments at XFEL or Synchrotron Sources
Pierre Aller and Allen M. Orville
12. From Tube to Structure: SPA Cryo-EM Workflow Using Apoferritin as an Example
Christoph A. Diebolder, Rebecca S. Dillard, and Ludovic Renault
13. Image Processing in Cryo-Electron Microscopy of Single Particles: The Power of Combining Methods
Carlos Oscar S. Sorzano, Amaya Jiménez-Moreno, David Maluenda, Erney Ramírez-Aportela, Marta Martínez, Ana Cuervo, Robert Melero, Jose Javier Conesa, Ruben Sánchez-García, David Strelak, Jiri Filipovic, Estrella Fernández-Giménez, Federico de Isidro-Gómez, David Herreros, Pablo Conesa, Laura del Caño, Yunior Fonseca, Jorge Jiménez de la Morena, Jose Ramon Macías, Patricia Losana, Roberto Marabini, and Jose-Maria Carazo
14. Setup and Troubleshooting of Volta Phase Plate Cryo-EM Data Collection
Ottilie von Loeffelholz and Bruno P. Klaholz
15. Cryo-Focused Ion Beam Lamella Preparation Protocol for In Situ Structural Biology
Jana Moravcová, Radka Dopitová, Matyáš Pinkas, and Jiří Nováček
16. Protein and Small Molecule Structure Determination by the Cryo-EM Method MicroED
Emma Danelius and Tamir Gonen
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.