Description
Insoluble Proteins, Softcover reprint of the original 1st ed. 2015
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1258
Language: EnglishSubject for Insoluble Proteins:
Support: Print on demand
Publication date: 12-2014
425 p. · 17.8x25.4 cm · Hardback
Description
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With insolubility proving to be one of the most crippling bottlenecks in the protein production and purification process, this volume serves to aid researchers working in the recombinant protein production field by describing a wide number of protocols and examples. Insoluble Proteins: Methods and Protocols includes chapters that describe not only the recombinant protein production in different expression systems but also different purification and characterization methods to finally obtain these difficult-to-obtain proteins. Beginning with protein production methods using both prokaryotic and eukaryotic expression systems, the book continues with purification protocols using insoluble proteins, the characterization of insoluble proteins, as well as a general overview of interesting applications of insoluble proteins. 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 tips on troubleshooting and avoiding known pitfalls.
Comprehensive and practical, Insoluble Proteins: Methods and Protocols aims to provide the scientific community with detailed and reliable state-of-the-art protocols that are used in order to successfully produce and purify recombinant proteins prone to aggregate.
1. General Introduction: Recombinant Protein Production and Purification of Insoluble Proteins
Neus Ferrer-Miralles, Paolo Saccardo, José Luis Corchero, Zhikun Xu, and Elena García-Fruitós
Part I: Recombinant Protein Production in Escherichia coli
2. Overcoming the Solubility Problem in E. coli: Available Approaches for Recombinant Protein Production
Agustín Correa and Pablo Oppezzo
3. Optimization of Culture Parameters and Novel Strategies to Improve Protein Solubility
Ranjana Arya, Jamal S.M. Sabir, Roop S. Bora, and Kulvinder S. Saini
4. Cleavable Self-Aggregating Tags (cSAT) for Protein Expression and Purification
Zhanglin Lin, Qing Zhao, Bihong Zhou, Lei Xing, and Wanghui Xu
5. Beyond the Cytoplasm of Escherichia coli: Localizing Recombinant Proteins Where You Want Them
Jason T. Boock, Dujduan Waraho-Zhmayev, Dario Mizrachi, and Matthew P. DeLisa
6. Characterization of Amyloid-Like Properties in Bacterial Intracellular Aggregates
Anna Villar-Pique, Susanna Navarro, and Salvador Ventura
Part II: Strategies to Produce Insoluble Proteins in Cell-Free Expression Systems
7. Co-Translational Stabilization of Insoluble Proteins in Cell-Free Expression Systems
Lei Kai, Erika Orbán, Erik Henrich, Davide Proverbio, Volker Dötsch, and Frank Bernhard
Part III: Recombinant Protein Production in Lactic Acid Bacteria (LAB)
8. Functional Expression of Plant Membrane Proteins in Lactococcus lactis
Sylvain Boutigny, Emeline Sautron, Annie Frelet-Barrand, Lucas Moyet, Daniel Salvi, Norbert Rolland, and Daphné Seigneurin-Berny
Part IV: Recombinant Protein Production in Yeast
9. High Cell-Density Expression System: Yeast Cells in a Phalanx Efficiently Produce a Certain Range of “Difficult-to-Express” Secretory Recombinant Proteins
Yasuaki Kawarasaki, Takeshi Kurose, and Keisuke Ito
Part V: Recombinant Protein Production in Insect Cells-Baculovirus
10. Insect Cells-Baculovirus System for the Production of Difficult to Express Proteins
Judit Osz-Papai, Laura Radu, Wassim Abdulrahman, Isabelle Kolb-Cheynel, Nathalie Troffer-Charlier, Catherine Birck, and Arnaud Poterszman
Part VI: Recombinant Protein Production in Mammalian Cells
11. Transient Expression in HEK 293 Cells: An Alternative to E. coli for the Production of Secreted and Intracellular Mammalian Proteins
Joanne E. Nettleship, Peter J. Watson, Nahid Rahman-Huq, Louise Fairall, Mareike G. Posner, Abhishek Upadhyay, Yamini Reddivari, Jonathan M.G. Chamberlain, Simon E. Kolstoe, Stefan Bagby, John W.R. Schwabe, and Raymond J. Owens
12. Recombinant Glycoprotein Production in Human Cell Lines
Kamilla Swiech, Marcela Cristina Corrêa de Freitas, Dimas Tadeu Covas, and Virgínia Picanço-Castro
Part VII: Recombinant Protein Production in Other Systems
13. Soluble Recombinant Protein Production in Pseudoaltermonas haloplanktis TAC125
Maria Giuliani, Ermenegilda Parrilli, Filomena Sannino, Gennaro Apuzzo, Gennaro Marino, and Maria Luisa Tutino
Part VIII: Insoluble Protein Purification
14. A Screening Methodology for Purifying Proteins with Aggregation Problems
Mario Lebendiker, Michal Maes, and Assaf Friedler
15. Solubilization and Refolding of Inclusion Body Proteins
Anupam Singh, Vaibhav Upadhyay, and Amulya K. Panda
16. Bacterial Inclusion Body Purification
Joaquin Seras-Franzoso, Spela Peternel, Olivia Cano-Garrido, Antonio Villaverde, and Elena García-Fruitós
17. Characterization of Intracellular Aggresomes by Fluorescent Microscopy
Lianwu Fu and Elizabeth Sztul
Part IX: Insoluble Protein Characterization
18. Dialysis: A Characterization Method of Aggregation Tendency
Mireia Pesarrodona, Ugutz Unzueta, and Esther Vázquez
19. Applications of Mass Spectrometry to the Study of Protein Aggregation
Sílvia Bronsoms and Sebastián A. Trejo
20. Insoluble Protein Assemblies Characterized by Fourier Transform Infrared Spectroscopy
Antonino Natalello and Silvia M. Doglia
21. Insoluble Protein Characterization by Circular Dichroism (CD) Spectroscopy and Nuclear Magnetic Resonance (NMR)
Shaveta Goyal, Haina Qin, Liangzhong Lim, and Jianxing Song
22. Methods for Characterization of Protein Aggregates
Witold Tatkiewicz, Elisa Elizondo, Evelyn Moreno, Cesar Díez-Gil, Nora Ventosa, Jaume Veciana, and Imma Ratera
23. Predicting the Solubility of Recombinant Proteins in Escherichia coli
Roger G. Harrison and Miguel J. Bagajewicz
Part X: Insoluble Protein Applications
24. Insoluble Protein Applications: The Use of Bacterial Inclusion Bodies as Biocatalysts
Eva Hrabárová, Lucia Achbergerová, and Jozef Nahálka
Includes cutting-edge methods and protocols involving insoluble proteins
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Includes supplementary material: sn.pub/extras