Insoluble Proteins: Methods and Protocols

418 Pages·2015·10.962 MB·English

by  Elena García-Fruitós (eds.)

Methods in
Molecular Biology 1258
Elena García-Fruitós Editor
Insoluble
Proteins
Methods and Protocols
M M B
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hat fi eld, Hertfordshire, AL10 9AB, UK
For further volumes:
http://www.springer.com/series/7651

Insoluble Proteins
Methods and Protocols
Edited by
Elena García-Fruitós
CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra (Barcelona), Spain;
Departament de Genètica i de Microbiologia, Institut de Biotecnologia i de Biomedicina,
Universitat Autònoma de Barcelona, Bellaterra (Barcelona), Spain
Editor
Elena G arcía-Fruitós
CIBER de Bioingeniería
Biomateriales y Nanomedicina (CIBER-BBN)
Bellaterra (Barcelona), Spain
Departament de Genètica i de Microbiologia
Institut de Biotecnologia i de Biomedicina
Universitat Autònoma de Barcelona
Bellaterra (Barcelona), Spain
ISSN 1064-3745 ISSN 1940-6029 (electronic)
ISBN 978-1-4939-2204-8 ISBN 978-1-4939-2205-5 (eBook)
DOI 10.1007/978-1-4939-2205-5
Springer New York Heidelberg Dordrecht London
Library of Congress Control Number: 2014955221
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Prefa ce
Since recombinant proteins are necessary for a wide range of applications for both biotechno-
logical and pharmaceutical industries, the interest in the recombinant protein production
fi eld has been growing exponentially in the last several years. In this context, although some
of these proteins are easily produced and purifi ed, many of them show important bottlenecks
in the production and purifi cation process with insolubility being one of the most important
ones. Thus, this volume of the Methods in Molecular Biology series aims to provide the scien-
tifi c 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. The main objective
of this book is to help those working in the recombinant protein production fi eld by describ-
ing a wide number of protocols and examples. The book is organized into 24 chapters that
describe not only the recombinant protein production in different expression systems but also
different purifi cation and characterization methods to fi nally obtain these diffi cult-to-obtain
proteins. Chapters 1 – 13 are focused on the description of protein production methods using
both prokaryotic and eukaryotic expression systems. Chapters 1 4 – 1 7 describe purifi cation
protocols using insoluble proteins, while Chapters 1 8 – 23 are useful to fi nd information
regarding the characterization of insoluble proteins. Finally, Chapter 2 4 aims to give a general
overview of interesting applications of insoluble proteins.
I would like to stress that this book has been written by a multidisciplinary team, which
adds value to its content since it has been analyzed from different points of view.
Finally, I would like to thank all the authors for their great job. The publication of this
book would not have been possible without the effort of all of them. I would also like to
thank Prof. John Walker for giving me the opportunity to edit this book and for his full
support through the whole process.
Bellaterra (Barcelona), Spain E lena García-Fruitós
v

Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x i
1 General Introduction: Recombinant Protein Production
and Purification of Insoluble Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Neus F errer-Miralles , P aolo Saccardo , José Luis Corchero ,
Zhikun Xu , and E lena G arcía-Fruitós
PART I RECOMBINANT PROTEIN PRODUCTION IN ESCHERICHIA COLI
2 Overcoming the Solubility Problem in E . coli : Available
Approaches for Recombinant Protein Production . . . . . . . . . . . . . . . . . . . . . . 27
Agustín C orrea and Pablo Oppezzo
3 O ptimization of Culture Parameters and Novel Strategies
to Improve Protein Solubility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Ranjana Arya , J amal S . M . S abir , Roop S . Bora ,
and Kulvinder S . S aini
4 C leavable Self-Aggregating Tags (cSAT) for Protein
Expression and Purification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Zhanglin Lin , Q ing Z hao , B ihong Z hou , Lei Xing , and Wanghui Xu
5 B eyond the Cytoplasm of E scherichia coli: Localizing
Recombinant Proteins Where You Want Them . . . . . . . . . . . . . . . . . . . . . . . . 79
Jason T. Boock , D ujduan W araho-Zhmayev , D ario Mizrachi ,
and Matthew P . D eLisa
6 Characterization of Amyloid-Like Properties in Bacterial
Intracellular Aggregates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
Anna Villar-Pique , Susanna Navarro , and S alvador V entura
PART II S TRATEGIES TO PRODUCE INSOLUBLE PROTEINS
IN CELL-FREE EXPRESSION SYSTEMS
7 Co-translational Stabilization of Insoluble Proteins
in Cell-F ree Expression Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 25
Lei K ai , E rika O rbán , E rik H enrich , Davide P roverbio ,
Volker D ötsch , and Frank B ernhard
PART III RECOMBINANT PROTEIN PRODUCTION
IN LACTIC ACID BACTERIA (LAB)
8 Functional Expression of Plant Membrane Proteins in L actococcus lactis . . . . . . 147
Sylvain B outigny , Emeline S autron , Annie F relet-Barrand ,
Lucas Moyet , D aniel S alvi , Norbert Rolland ,
and Daphné Seigneurin-Berny
vii
viii Contents
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. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
Yasuaki Kawarasaki , T akeshi Kurose , and Keisuke I to
PART V RECOMBINANT PROTEIN PRODUCTION
IN INSECT CELLS-BACULOVIRUS
10 Insect Cells–Baculovirus System for the Production
of Difficult to Express Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 81
Judit O sz-Papai , Laura R adu , W assim A bdulrahman ,
Isabelle K olb-C heynel , Nathalie T roffer-Charlier , C atherine 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 . . . . . . . 209
Joanne E. Nettleship , Peter J. W atson , Nahid Rahman-Huq ,
Louise Fairall , M areike G. P osner , Abhishek Upadhyay ,
Yamini Reddivari , J onathan M . G . Chamberlain , Simon E . Kolstoe ,
Stefan B agby , J ohn W. R . S chwabe , and Raymond J. Owens
12 R ecombinant Glycoprotein Production in Human Cell Lines . . . . . . . . . . . . . 2 23
Kamilla S wiech , M arcela C ristina Corrêa de Freitas ,
Dimas Tadeu C ovas , and Virgínia Picanço-Castro
PART VII RECOMBINANT PROTEIN PRODUCTION IN OTHER SYSTEMS
13 Soluble Recombinant Protein Production in P seudoalteromonas
haloplanktis TAC125. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 43
Maria Giuliani , E rmenegilda P arrilli , F ilomena S annino ,
Gennaro Apuzzo , G ennaro M arino , and Maria L uisa Tutino
PART VIII INSOLUBLE PROTEIN PURIFICATION
14 A Screening Methodology for Purifying Proteins with Aggregation
Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 61
Mario Lebendiker , M ichal Maes , and Assaf Friedler
15 S olubilization and Refolding of Inclusion Body Proteins. . . . . . . . . . . . . . . . . 2 83
Anupam Singh , V aibhav Upadhyay , and A mulya K. Panda
16 Bacterial Inclusion Body Purification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 293
Joaquin S eras-F ranzoso , Spela P eternel , Olivia Cano-Garrido ,
Antonio V illaverde , and Elena G arcía-Fruitós
17 C haracterization of Intracellular Aggresomes by Fluorescent Microscopy. . . . . 3 07
Lianwu Fu and E lizabeth S ztul
Contents ix
PART IX INSOLUBLE PROTEIN CHARACTERIZATION
18 Dialysis: A Characterization Method of Aggregation Tendency . . . . . . . . . . . . 3 21
Mireia Pesarrodona , U gutz Unzueta , and E sther V ázquez
19 A pplications of Mass Spectrometry to the Study of Protein Aggregation . . . . . 3 31
Sílvia B ronsoms and Sebastián A. T rejo
20 I nsoluble Protein Assemblies Characterized by Fourier
Transform Infrared Spectroscopy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 347
Antonino Natalello and Silvia M . D oglia
21 Insoluble Protein Characterization by Circular Dichroism (CD)
Spectroscopy and Nuclear Magnetic Resonance (NMR) . . . . . . . . . . . . . . . . . 3 71
Shaveta Goyal , H aina Q in , Liangzhong Lim , and Jianxing S ong
22 Methods for Characterization of Protein Aggregates. . . . . . . . . . . . . . . . . . . . 3 87
Witold T atkiewicz , Elisa E lizondo , E velyn Moreno , C esar D íez-Gil ,
N ora Ventosa , Jaume V eciana , and Imma R atera
23 P redicting the Solubility of Recombinant Proteins in E scherichia coli . . . . . . . . 403
Roger G . H arrison and Miguel J. B agajewicz
PART X INSOLUBLE PROTEIN APPLICATIONS
24 Insoluble Protein Applications: The Use of Bacterial Inclusion
Bodies as Biocatalysts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 11
Eva Hrabárová , Lucia A chbergerová , and Jozef N ahálka
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 23