Membrane proteins: production and functional characterization:
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Amsterdam [u.a.]
Elsevier
2015
|
| Edition: | 1. ed. |
| Series: | Methods in enzymology
556 |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Academic Press is an imprint of Elsevier |
| Physical Description: | XXIV, 651 S., [8] Bl. Ill., graph. Darst. |
| ISBN: | 9780128015216 |
Staff View
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Record in the Search Index
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| adam_text | CONTENTS
Contributors xv
Preface XX/ / /
Section I
Recombinant Expression of Membrane Proteins
1. Engineering Escherichia coli for Functional Expression
of Membrane Proteins 3
Franz Y. Ho and Bert Poolman
1. Introduction 4
2. Preparation of Erythromycin-Sensitive E. coli Strain 8
3. Preparation of Expression Plasmid 10
4. Selecting Cells for Better Expression 14
5. Characterizing Evolved Strains 15
6. Summary 17
Acknowledgments 18
References 18
2. ACEMBLing a Multiprotein Transmembrane Complex: The
Functional SecYEG-SecDF-YajC-YidC Holotranslocon Protein
Secretase/lnsertase 23
Joanna Komar, Mathieu Botte, Ian Collinson, Christiane Schaffitzel,
and Imre Berger
1. Introduction 24
2. ACEMBLing the HTL Multiprotein Complex 27
3. Purifying the HTL 28
4. HTL Integrity and Activity 32
5. Discussion and Conclusions 43
Acknowledgments 46
References 46
3. Expression and Purification of OsVDAC4 51
Ashwini Godbole, Ashvini K. Dubey, Rohan Mitra, Palakolanu S. Reddy,
and M.K. Mathew
1. Introduction 52
2. Equipment 53
v
VI
Contents
3. Materials 53
4. Protocols 57
Acknowledgments 74
References 74
4. Membrane Protein Expression in Lactococcus lactis 77
Martin S. King, Christoph Boes, and Edmund R.S. Kunji
1. Introduction 78
2. Equipment and Materials 84
3. Buffers and Media 85
4. Protocol 87
5. Step 1: Cloning the Target Gene into pNZ8048 and
Transformation into /_. lactis 89
6. Step 2: Growth of L lactis, Expression of the Target Protein and
Preparation of Membrane Vesicles 94
Acknowledgment 96
References 96
5. An Unconventional Anaerobic Membrane Protein
Production System Based on Wolinella succinogenes 99
Michael Lafontaine and C. Roy D. Lancaster
1. Introduction 100
2. Methods 106
3. Purification of Proteins Expressed in IV. succinogenes 111
Acknowledgments 118
References 119
6. Membrane Protein Expression and Analysis in Yeast 123
Katrien Claes, Mouna Guerfal, and Nico Callewaert
V Theory 124
2. Equipment 125
3. Materials 126
4. Protocol 128
5. Step 1: Transformation of Y. lipolytica 129
6. Step 2: Small-Scale Membrane Protein Expression 132
7. Step 3: Membrane Protein Preparation 133
8. Step 4: Membrane Protein Analysis: Expression Levels 134
9. Step 5: Membrane Protein Analysis: Functionality 135
References 139
Further reading 140
Contents vii
7. Heterologous Expression of G-Protein-Coupled
Receptors in Yeast 141
Nicolas Bertheleme, Shweta Singh, Simon Dowell, and Bernadette Byrne
1. Introduction 142
2. Pichia posions 142
3. Soccharomyces cerevisioe 144
4. Which System to Choose? 146
5. Generating P. postons Expression Plasmids 146
6. Generating and Screening the P. postons Expression Clones 147
7. Some Considerations 150
8. Generating S. cerevisioe Expression Clones 151
9. In-Gel Fluorescence 152
10. Confoca . Microscopy 153
11. Assessment of Protein Expression Levels 155
12. Yeast Cell Growth Assay 156
13. Optimization of the Assay Conditions 157
14. Outcome of the Functional Analysis 158
15. Conclusions 159
Acknowlecgments 159
References 160
8. Recombinant G Protein-Coupled Receptor Expression in
Soccharomyces cerevisioe for Protein Characterization 165
Kory M. Blocker, Zachary T. Britton, Andrea N. Naranjo, Patrick M. McNeely,
Carissa L. Young, and Anne S. Robinson
1. Introduction 166
2. Plasmid Design 167
3. Characterization 169
4. Conclusion 179
Acknowledgments 180
References 180
9. Baculovirus-Mediated Expression of GPCRs in Insect Cells 185
Tuulia Saarenpää, Veli-Pekka Jaakola, and Adrian Goldman
1. Introduction 186
2. Cell Maintenance 188
3. Recombinant Bacmid DNA 191
4. Transfection, Amplification, and Storage of Virus 208
5. Protein Expression 213
References 216
Contents
viii
10. Expression of Membrane Proteins in the Eyes of Transgenic
Drosophila melanogaster 219
Yvonne Hackmann, Lisa Joedicke, Valerie Panneels, and Irmgard Sinning
1. introduction 220
2. Protein Expression in Transgenic Drosophila melanogaster 221
3. Protocol 224
4. Case Study: Signal Peptide Peptidase 236
5. Conclusions and Outlook 236
References 238
11. Rapid Method to Express and Purify Human Membrane
Protein Using the Xenopus Oocyte System for Functional
and Low-Resolution Structural Analysis 241
Benjamin Clémengon, Michael Fine, Philipp Schneider, and
Matthias A. Hediger
1. Introduction 242
2. Subcloning in pMJB08 and cRNA Production 243
3. Frog Surgery, Oocytes Preparation and Injection 244
4. Determination of Maximal Level Expression 251
5. Membrane Preparation and Detergent Screening 253
6. Membrane Protein Purification: Ion Metal Affinity Chromatography 256
7. Posttranslational Verification Using Deglycosylation 258
8. Determination of Surface Membrane Expression Using Surface
Biotinylation 259
9. Functional Verification of Membrane Proteins in the Oocyte 260
10. Grids Preparation and Negative Staining 261
11. Image Processing 261
12. Conclusion 264
Acknowledgments 264
References 264
1 2. Expression of G Protein Coupled Receptors in Mammalian Cells 267
Raja Chakraborty, Bing Xu, Rajinder P. Bhuilar, and Prashen Chelikani
1. Introduction 268
2. Materials and Supplies 270
3. Procedure 273
Acknowledgments 279
References 280
Contents ix
1 3. Construction of Stable Mammalian Cell Lines for Inducible
Expression of G Protein-Coupled Receptors 283
Chikwado A. Opefi, Dale Tranter, Steven 0. Smith, and Philip J. Reeves
1. Introduction 284
2. Construction of Stable HEK293S Cell Lines Containing Integrated Genes
Regulated by Tetracycline 286
3. Growth of Tetracycline-Inducible Stable Cell Lines in
Suspension Culture 299
4. Purification of Membrane Proteins Tagged with the Antirhodopsin
Rho-1 D4 Antibody 301
Acknowledgments 304
References 304
14. Rapid and Facile Recombinant Expression of Bovine
Rhodopsin in HEK293S GnTI՜ Cells Using a PiggyBac
Inducible System 307
Lydia N. Caro, Zhijie Li, Aidin R. Balo, Ned Van Bps, James M. Rini,
and Oliver P. Ernst
1. Theory 308
2. Protocol 310
3. Step 1: Subcloning of Wild-Type and Mutant Opsin cDNA into
Plasmid, PB-T-PAF 313
4. Step 2 Transfection, Transient Expression, and Generation of
Stable Bulk Cell Cultures 317
5. Step 3: Scale-Up of Bulk Cell Cultures in Roller Bottles 320
6. Step 4: Induction and Cell Harvesting 321
7. Step 5: Rhodopsin Purification and Spin Labeling 323
8. Conclusion 326
Acknowledgments 328
References 328
15.Semliki Forest Virus Based Expression of Recombinant GPCRs 331
Kenneth tundstrom
1. Introduction 332
2. $FV Expression System 333
3. Expression of GPCRs 339
4. Summary 346
References 348
X
Contents
16. Screening for Lipid Requirements of Membrane Proteins by
Combining Cell-Free Expression with Nanodiscs 351
Erik Henrich, Volker Dotsch, and Frank Bernhard
1. Introduction 352
2. General Considerations 353
3. Protocol: CF Lysate Preparation 354
4. Protocol: Preparation of NDs and Liposomes 356
5. Protocol: Optimizing the Efficiency of (IF Expression 359
6. Protocol: CF Expression Reactions in the L-CF Mode 360
7. Protocol: I ipid Modulation of Membrane Protein Activity:
Ihe Example of MraY Translocates 363
8. Protocol: Purification of CF-Fxpressed Membrane Proteins 365
9. Conclusion and Outlook 367
Acknowledgments 36/
References 367
Section II
Reconstitution and Functional Characterization
of Membrane Proteins
17. Liposome Reconstitution and Transport Assay for Recombinant
Transporters 373
Zachary Lee Johnson and Seok-Yong Lee
1. Introduction 373
2. Reconsitution of Protein into Liposomes 375
3. Assaying Transport 378
4. Summary 381
Acknowledgment 382
References 382
18. Lipid Reconstitution and Recording of Recombinant
Ion Channels 385
Randy B. Stockbridge and Ming-Feng Tsai
1. Introduction 386
2. Liposome Reconstitution of Ion Channels 386
3. Liposome Flux Assays 391
4. Electrical Recording of Reconstituted Protein 395
5. Summary 402
Acknowledgments 402
References 402
contents Xi
19. Reconstitution of Membrane Proteins: A GPCR as an Example 405
Alan D. Goddard, Patricia M. Dijkman, Roslin J. Adamson,
Rosana inacio dos Reis, and Anthony Watts
1. Introduction 406
2. Production of Liposomes 406
3. Reconstitution into Liposomes 409
4. Reconstitution into Nanodiscs 413
5. Reconstitution into Lipodisqs 416
6. Determination of Lipid-to-Protein Ratio 418
7. Concentration Methods 421
8. Summary 421
Acknowledgments 422
References 4 22
20. Ion Channel Reporter for Monitoring the Activity of
Engineered GPCRs 425
Christophe J. Moreau, Katarzyna Niescierowicz, l ydia N. Caro,
Jean Revilloud, and Michel Vivaudou
1. Introduction 426
2. ICCR Design 440
3. Functional Characterization 446
4. Current Advantages and Limitations of the ICCR Technology 451
Acknowledgments 452
References 452
21. Methods for Labeling Skeletal Muscle Ion Channels
Site-Specifically with Fluorophores Suitable for FRET-Based
Structural Analysis 455
Mohana Mahalingam and James D. Fessenden
1. Introduction 456
2. Fluorescent Protein (FP) Fusions 458
3. Labeling of Tetracysteine (Tc) Tags by Bïarsenical Fluorophores 4-61
4. Labeling of Poly-Histidine Tags by Fluorescent NTA Conjugates 464
5. Three-Way FRET Using a Combination of Site-Specific Labeling Methods 469
6. Summary 471
Acknowledgments 471
References 471
xi i Contents
22. Structural and Functional Studies of NirC from Salmonella
typhimurium 475
Adriana Rycovska Blume; Wei Lu, Susana Andrade, Klaus Fendler,
and Oliver Einsle
1. Introduction 476
2. Production and Isolation of SfNirC 477
3. Structural Analysis of SfNirC 480
4. Functional Assays for SfNirC 484
5. Conclusions 494
Acknowledgments 495
References 495
23. Surface Plasmon Resonance Analysis of
Seven-Transmembrane Receptors 499
Tonia Aristotelous, Andrew L. Hopkins, and Iva Navratilova
1. introduction 500
2. Surface Plasmon Resonance 501
3. 5PR Approaches to FBDD 503
4. SPR Applications for GPCRs SÜ7
5. SPR Applications for Allosteric Compounds 514
6. SPR Fragment Screening of Thermostabilized GPCRs 516
7. 5PR Fragment Screening of Fully Functional GPCRs 517
8. Confirmation of SPR Fragment Hits 521
9. Conclusion 522
References 523
24. Cross-linking Strategies to Study Peptide Ligand-Receptor
Interactions 527
Jeffrey M. Becker and Fred Naider
1. Introduction 528
2. Photochemical Cross-linking 529
3. Chemical Cross-linking 534
4. Cross-Linking by Unnatural Amino Acid Replacement into the Receptor 537
5. Identifying the Cross-link Site 539
6. Concluding Remarks 543
Acknowledgments 544
References 544
Contents
XIII
25. From Recombinant Expression to Crystals: A Step-by-Step
Guide to GPCR Crystallography 549
Arun K. Shukla, Punita Kumari, Eshan Ghosh, and Kumari Nidhi
1. Introduction 550
2. Target Selection, Construct Design, and Protein Engineering 550
3. Recombinant Expression and Selection of Host System 553
4. Functional Solubilization and Homogenous Purification 554
5. Approaches for Crystallogenesis and Optimization 556
6. Conclusion and Future Perspectives 558
Acknowledgments 558
References 558
26. Structure Based Biophysical Analysis of the Interaction of
Rhodopsin with G Protein and Arrestin 563
Martha E. Sommer, Matthias Elgeti, Peter W. Hildebrand, Michal Szczepek,
Klaus Peter Hofmann, and Patrick Scheerer
1. ntroduCtion 564
2. Preparation of Proteins and Peptides 567
3. Protein X-ray Crystallography 572
4. UVA/is Absorption Spectroscopy: Extra Meta II 578
5. Site-Directed Fluorescence Spectroscopy 582
6. FTIR Difference Spectroscopy 589
7. Molecular Dynamics Simulations 595
Acknowledgments 601
References 602
Author Index 609
Subject Index 639
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| spelling | Membrane proteins: production and functional characterization ed. by Arun K. Shukla 1. ed. Amsterdam [u.a.] Elsevier 2015 XXIV, 651 S., [8] Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Methods in enzymology 556 Academic Press is an imprint of Elsevier Membranproteine (DE-588)4130026-9 gnd rswk-swf (DE-588)4143413-4 Aufsatzsammlung gnd-content Membranproteine (DE-588)4130026-9 s DE-604 Shukla, Arun K. 1981- Sonstige (DE-588)131837397 oth Methods in enzymology 556 (DE-604)BV000000938 556 Digitalisierung UB Regensburg - ADAM Catalogue Enrichment application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027966524&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Membrane proteins: production and functional characterization Methods in enzymology Membranproteine (DE-588)4130026-9 gnd |
| subject_GND | (DE-588)4130026-9 (DE-588)4143413-4 |
| title | Membrane proteins: production and functional characterization |
| title_auth | Membrane proteins: production and functional characterization |
| title_exact_search | Membrane proteins: production and functional characterization |
| title_full | Membrane proteins: production and functional characterization ed. by Arun K. Shukla |
| title_fullStr | Membrane proteins: production and functional characterization ed. by Arun K. Shukla |
| title_full_unstemmed | Membrane proteins: production and functional characterization ed. by Arun K. Shukla |
| title_short | Membrane proteins: production and functional characterization |
| title_sort | membrane proteins production and functional characterization |
| topic | Membranproteine (DE-588)4130026-9 gnd |
| topic_facet | Membranproteine Aufsatzsammlung |
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