Membrane Protein Structure and Function Characterization, 1st ed. 2017
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1635

In this present volume, different approaches are detailed to produce membrane proteins, purify them, study their function, determine their structure, and model them in membrane. Since every membrane protein behaves mostly in a unique way /fashion, knowledge of guidelines and tricks may help to increase chances to express, purify and characterize a peculiar membrane protein. Production of correctly folded protein remains a challenge. Moreover, getting a functional and stable protein requires to optimize membrane mimicking environments that can be detergent or artificial membranes. In some cases, the finding of the correct ligand which will stabilize the desired conformation is needed. In other cases, stabilization can be obtained using specific antibodies. This volume also presents different techniques to analyze the functional status of membrane proteins. Written in the highly successful Methods in Molecular Biology series format, chapters in Membrane Protein Structure and Function Characterization: Methods and Protocols providedifferent techniques to analyze the functional and structural status of membrane proteins.  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.

Authoritative and practical, Membrane Protein Structure and Function Characterization: Methods and Protocols aims to ensure successful results in the further study of this vital field. 

Lucile Senicourt, Soria Iatmanen-Harbi, Claude Hattab, Mariano A Ostuni, Marie-France Giraud, and Jean-Jacques Lacapere

2. Functional Assembly of Soluble and Membrane Recombinant Proteins of Mammalian NADPH Oxidase Complex

Hajer Souabni, Aymen Ezzine, Tania Bizouarn, and Laura Baciou

3. Direct Extraction and Purification of Recombinant Membrane Proteins from Pichia pastoris protoplasts

Lucie Hartmann, Estelle Metzger, Noémie Ottelard, and Renaud Wagner

4. Cell-free Expression for the Study of Hydrophobic Proteins: The Example of Yeast ATP-synthase subunits

Isabelle Larieu, James Tolchard, Corinne Sanchez, Edmond Yazo Kone, Alexandre Barras, Claire Stines-Chaumeil, Benoit Odaert,  and  Marie-France Giraud

5. Wheat Germ Cell Free Overexpression for the Production of Membrane Proteins

Marie-Laure Faugeron, Aurelie Badillo, Francois Penin, and Anja Bocjmann

6. Methyl-specific Isotope Labelling Strategies for NMR Studies of Membrane Proteins

Vilius Kurauskas, Paul Schanda, and Remy Sounier

7. Labeling of Membrane Complexes for Electron Microscopy

Francesca Gubellini and Rémi Fronzes

8. Expression, Biochemistry, and Stabilization with Camel Antibodies of Membrane Proteins Case Study of The Mouse 5-HT Receptor

Ghérici Hassaine, Cédric Deluz, Luigino Grasso, Romain Wyss, Ruud Hovius, Henning Stahlberg, Takashi Tomizaki, Aline Desmyter, Christophe Moreau, Lucie Peclinovska, Sonja Minniberger, Lamia Mebarki, Xiao-Dan Li, Horst Vogel, and Hugues Nury

9. Characterization of New Detergent and Detergent Mimetics by Scattering Techniques for Membrane Protein Crystallization

Francoise Bonnete and Patrick J Loll

10. Secondary Structure Determination by Means of ATR-FTIR Spectroscopy

Batoul Srour, Stefan Bruechert, Susana Andrade, and Petra Hellwig

11. Native Mass Spectroscopy for the Characterization of Structure and Interactions of Membrane Proteins

Jeroen F Van Dyck, Albert Konijnenberg, and Frank Sobott

12. Mass Spectroscopy of Mitochondrial Membrane Proteins Complexes                        

Luc NEGRONI, Michel ZIVY, and Claire LEMAIRE

13. Functional Studies on Membrane Proteins by Means of H/D Exchange in Infrared: Structural Changes in Na⁺NQR from V cholerae in the Presence of Lipids

Yashvin Neehaul, Sebastien Kriegel, Blanca Barquera, and Petra Hellwig

14. Reconstitution of Membrane Protein in Liposomes

Alice Verchere, Isabelle Broutin, and Martin Picard

15. Ion Channels as Reporters of Membrane Receptor Function: Automated Analysis in Xenopus Oocytes

Michel Vivaudou, Zlatomir Todorov, Gina Catalina Reyes-Mejia, and Christophe Moreau

16. The CRACAM Robot: Two Dimensional Crystallization of Membrane Protein

Philippe Rosier, Frederic Gelebart, Nicolas Dumesnil, Gauthier Esnot, Manuela Dezi, Marc Morand, and Catherine Venien-Bryan

17. Reconstitution of Membrane Proteins into Nanodiscs for Single Particle Electron Microscopy

Laetitia Daury, Jean-Christophe Taveau, Dimitri Salvador, Marie Glavier, and Olivier Lambert

18. Solid-state NMR of Membrane Protein Reconstituted in Proteoliposomes, the Case of TSPO

Lucile Senicourt, Luminita Duma, Vassilios Papadopoulos, and Jean-Jacques Lacapere

19. Sample Preparation for Membrane Protein Structural Studies by Solid-State NMR

Denis Lacabanne, Britta Kunert, Carole Gardiennet, Beat H Meier, and Anja Bockmann

20. Simulation of Ligand Binding To Membrane Proteins

Samuel Murail

21. Molecular modelling of Transporters: From Low Resolution Cryo-Electron Microscopy Map to Conformational Exploration. The Example of TSPO Molecular Dynamic of Membrane Protein in Different Environments

Aurore Vaitinadapoule and Catherine Etchebest

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts

Includes supplementary material: sn.pub/extras

Includes supplementary material: sn.pub/extras