Advanced Technologies for Protein Complex Production and Characterization

ISBN-13: 9783319272146 / Angielski / Twarda / 2016 / 382 str.

M. Cristina Vega

Opis

This book presents advanced expression technologies for the production of protein complexes. Since complexes lie at the heart of modern biology, the expression, purification, and characterization of large amounts of high-quality protein complexes is crucial for the fields of biomedicine, biotechnology, and structural biology. From co-expression in E. coli, yeast, mammalian and insect cells to complex reconstitution from individual subunits, this book offers useful insights and guidance for successful protein expressionists. Across several sections readers will discover existing opportunities for the production of protein complexes in bacterial systems (including membrane proteins and cell-free co-expression), methylotrophic and non-methylotrophic yeasts, protozoa (Leishmania terantolae and Dictyostelium discoideum), baculovirus-infected insect cells, mammalian cells, plants and algae. Complex reconstitution from individually purified subunits or subcomplexes is discussed as a complementary strategy. A last section introduces briefly some of the biophysical and structural characterization techniques for macromolecular complexes using state-of-the-art solution scattering and nuclear magnetic resonance. This work is a guided tour over some of the most powerful and successful protein expression technologies, with a focus on co-expression and high-throughput applications. It is addressed to everyone interested in the production and characterization of macromolecular complexes, from university students who want an accessible description of the major co-expression systems to researchers in biomedicine and the life sciences seeking for an up-to-date survey of available technologies.

Spis treści

Preface. Complex Biologics: Engineering the Tools and the Hosts. M. Cristina Vega, Francisco J. Fernández and Imre Berger

Section 1: Introduction

1. Protein complex production from the drug discovery standpoint; Ismail Moarefi     

<2. Choose a suitable expression host: a survey of available protein production platforms; Francisco J. Fernández and M. Cristina Vega

Section 2: Prokaryotic expression hosts

3. ACEMBL Tool-Kits for High-Throughput Multigene Delivery and Expression in Prokaryotic and Eukaryotic Hosts; Yan Nie, Maxime Chaillet, Christian Becke, Matthias Haffke, Martin Pelosse, Daniel Fitzgerald, Ian Collinson, Christiane Schaffitzel and Imre Berger

4. Complex reconstitution and characterization by combining co-expression techniques in Escherichia coli with high-throughput; Renaud Vincentelli and Christoph Romier

5. Membrane protein production in E. coli for applications in drug discovery; Harm Jan (Arjan) Snijder and Jonna Hakulinen

6. Cell-free Synthesis of Macromolecular Complexes; Matthieu Botte, Aurélien Deniaud and Christiane Schaffitzel                                                                                         

7. A Bacillus megaterium system for the production of recombinant proteins and protein complexes; Rebekka Biedendieck

8. Protein complex production in alternative prokaryotic hosts; Sara Gómez, Miguel López-Estepa, Francisco J. Fernández and M. Cristina Vega                              

Section 3: Lower Eukaryotic expression hosts

9. Production of Protein Complexes in Non-methylotrophic and Methylotrophic Yeasts; Francisco J. Fernandez, Miguel López-Estepa, Javier Querol-García and M. Cristina Vega

10. Leishmania tarentolae for the Production of Multi-Subunit Complexes; Tomoaki Niimi

11. Alternative Eukaryotic Expression Systems for the Production of Proteins and Protein Complexes; Sara Gómez, Miguel López-Estepa, Francisco J. Fernández, Teresa Suárez and M. Cristina Vega

Section 4: Higher Eukaryotic expression hosts

12. Fundamentals of baculovirus expression and applications; Tom A. Kost and Christopher W. Kemp

13. The MultiBac Baculovirus / Insect Cell Expression Vector System for Producing Complex Protein Biologics; Duygu Sari, Kapil Gupta, Deepak Balaji Govinda Raj, Alice Aubert, Petra Dmcova, Frederic Garzoni, Daniel Fitzgerald and Imre Berger  

14. Fundamentals of expression in mammalian cells; Michael R. Dyson 

15. Assembling multi-subunit complexes using mammalian expression; Bahar Baser and Joop van den Heuvel

Section 5: Plant expression

16. Microalgae as solar-powered protein factories; Franziska Hempel and Uwe G. Maier

17. Strategies and methodologies for the co-expression of multiple proteins in plants; Albert Ferrer, Montserrat Arró, David Manzano and Teresa Altabella                

18. Transient expression systems in plants—potentialities and constraints; Tomas Canto         

Section 6: Complex reconstitution

19. Complex reconstitution from individual protein modules; Jérome Basquin, Michael Taschner and Esben Lorentzen                                                                             

20. Structural reconstruction of protein-protein complexes involved in intracellular signaling; Klára Kirsch, Péter Sok, and Attila Remény                                                           

Section 7: Biophysical methods to assess protein complexes

21. The use of small-angle scattering for the characterization of multi subunit complexes; Adam Round

22. Application of nuclear magnetic resonance and hybrid methods to structure determination of complex systems. Filippo Prischi and Annalisa Pastore

O autorze

Dr. M. Cristina Vega is a group leader at the Chemical and Physical Biology Department of the Center for Biological Research (CIB-CSIC) in Madrid, Spain. With substantial experience in macromolecular X-ray crystallography of protein complexes with biomedical relevance, she designs eukaryotic, yeast-based expression platforms tailored to protein complexes with a focus on cost-efficiency and improved performance.