Insoluble Proteins: Methods and Protocols

ISBN-13: 9781071618585 / Angielski / Twarda / 2022 / 600 str.

Elena Garcia Fruitós; Anna Arís Giralt

Spis treści

1. General Introduction: Recombinant protein production and purification of insoluble proteins

Neus Ferrer-Miralles, Paolo Saccardo, José Luis Corchero, and Elena García-Fruitós

 

Part I: Prokaryotic Expression Systems

2. Overcoming the solubility problem in E. coli: available approaches for recombinant protein production

Claudia Ortega, Pablo Oppezzo, and Agustín Correa

 

3. A3D 2.0 update for the prediction and optimization of protein solubility

Jordi Pujols, Valentín Iglesias, Jaime Santos, Aleksander Kuriata, Sebastian Kmiecik, and Salvador Ventura

 

4. Tailoring codon usage to the underlying biology for protein expression optimization

Zahra Alirezaeizanjani, Jan-Hendrik Trösemeier, Christel Kamp, and Sophia Rudorf

 

5. Protein Expression Optimization Strategies in E. coli: A Tailored Approach in Strain Selection and Parallelizing Expression Conditions

Shyn Ric Tang, Balaji Somasundaram, and Linda H.L. Lua

 

6. A novel approach for production of aggregation prone proteins using the spidroin-derived NT* tag

Nina Kronqvist, Anna Rising, and Jan Johansson

 

7. Cleavable Self-Aggregating Tags (cSAT) for Therapeutic Peptide Expression and Purification

Xiaofeng Yang, Zhanglin Lin, and Yanyun Jing

 

8. Use Intein Cleavable Polyhydroxyalkanoate Synthase Fusions to Improve Protein Solubility

Shuxiong Chen and Bernd Rehm

 

9. Introduction of a hexalysine (6K) tag can protect from N-terminal cleavage and increase yield of recombinant proteins expressed in the periplasm of E. coli

Vasiliki Paraskevopoulou, Mohammed Alissa, Naim Hage, and Franco H. Falcone

 

10. Two-tiered selection and screening strategy to increase functional enzyme production in Escherichia coli

Jason T. Boock, May Taw, Brian C. King, Robert J. Conrado, Donna M. Gibson, and Matthew P. DeLisa

11. Method for production of cysteine-rich proteins in Lactococcus lactis expression system

Susheel K. Singh and Vandana Singh

 

12. Heterologous protein production in Lactobacillus (plantarum) using pSIP vectors

Geir Mathiesen, Lars Axelsson, Vincent GH Eijsink

 

13. Soluble recombinant protein production in Pseudoaltermonas haloplanktis TAC125: the case study of the full-length human CDKL5 protein

Marzia Calvanese, Andrea Colarusso, Concetta Lauro, Ermenegilda Parrilli and Maria Luisa Tutino

 

14. A protocol to enhance soluble protein expression in the cytoplasm of Bacillus subtilis

Hoang D Nguyen and Trang TP Phan

 

15. Applications of cell-free synthesized membrane protein precipitates

Julija Mezhyrova, Karsten Mörs, Clemens Glaubitz, Volker Dötsch, and Frank Bernhard

 

Part II: Eukaryotic Expression Systems

16. 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, Sayaka Ohashi, Runa Watabe, Mizuki Tanaka, and Keisuke Ito

 

17. Insect Cells-Baculovirus system for the production of difficult to express proteins: from expression screening for soluble constructs to protein quality control

Simon Pichard, Nathalie Troffer-Charlier, Isabelle Kolb-Cheynel, Pierre Poussin-Courmontagne, Wassim Abdulrahman, Catherine Birck, Vincent Cura, and Arnaud Poterszman

 

18. Transient gene expression in human Expi293 cells

Haroldo Cid da Silva Junior

 

19. Recombinant Expression of Complex Proteins in Human Cells Lines

Aline de Sousa Bomfim, Bruna Samhan Archangelo, Aline Sanches Pereira, Elisa Maria de Sousa Russo

 

Part III: Insoluble Protein Purification

20. Optimizing chaperone removal strategy from over-expressed recombinant proteins: GNE, a case study

Shweta Sharma, Roop Singh Bora, Kulvinder Singh Saini and Ranjana Arya

 

21. Intrinsically disordered proteins (IDP): purification under denaturing conditions

Mireia Pesarrodona, Isabel Latorre, and Xavier Salvatella

 

22. Solubilization and Refolding of Inclusion Body Proteins

Priyank Singhvi and Amulya K. Panda

 

23. Non-denaturing solubilization of Inclusion Bodies from Lactic Acid Bacteria

Laia Gifre-Renom, Ricardo Baltà-Foix, Anna Arís, and Elena Garcia-Fruitós

 

24. Purification of Inclusion Bodies Produced in Bacteria and Yeast

Joaquin Seras- Franzoso, Olivia Cano-Garrido, Spela Peternel, Anna Arís, and Elena García-Fruitós

 

25. Eukaryotic aggresomes: protocols and tips for their production, purification and handling

Rosa Mendoza, Neus Ferrer-Miralles, and José Luis Corchero

 

Part IV: Insoluble Protein Characterization

26. Characterization of the conformational properties of soluble and insoluble proteins by Fourier transform infrared spectroscopy

Diletta Ami and Antonino Natalello

 

27. Characterizing Soluble Protein Aggregates using Native Mass Spectrometry Coupled with Temperature-control Electrospray Ionization and Size-exclusion Chromatography

Khaja Muneeruddin, Igor A. Kaltashov, Guanbo Wang

28. Quality control of proteins solubilized from inclusion bodies

Julieta M. Sanchez, Jose Vicente Carratalá, Laia Gifre-Renom, Anna Arís, Elena Garcia-Fruitós, Neus Ferrer-Miralles

 

29. Methods for the Characterization of Protein Aggregates

Marc Martínez-Miguel, Witold Tatkiewicz, Mariana Köber, Nora Ventosa, Jaume Veciana, Judith Guasch and Imma Ratera

 

Part V: Insoluble Protein Applications

30. Insoluble Protein Applications: The Use of Bacterial Inclusion Bodies as Biocatalysts

Romana Köszagová, Eva Hrabárová, Lucia Achbergerová, and Jozef Nahálka

 

31. Methods for Processing Protein Aggregates into Surfaces

Marc Martínez-Miguel, Witold Tatkiewicz, Mariana Köber, Nora Ventosa, Jaume Veciana, Judith Guasch, and Imma Ratera

Blurb

This volume serves to aid researchers working in the recombinant protein production field by describing a wide number of protocols and examples. Chapters describe recombinant protein production in different expression systems, prokaryotic and eukaryotic expression systems, purification protocols, characterization of insoluble proteins and a general overview of interesting applications of insoluble proteins. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

 

Authoritative and cutting-edge, Insoluble Proteins: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field.