Membrane Protein Structure and Function Characterization

Methods and Protocols
 HC runder Rücken kaschiert
ISBN-13:
9781493971497
Veröffentl:
2017
Einband:
HC runder Rücken kaschiert
Erscheinungsdatum:
30.07.2017
Seiten:
432
Autor:
Jean-Jacques Lacapere
Gewicht:
1005 g
Format:
260x183x29 mm
Serie:
1635, Methods in Molecular Biology
Sprache:
Englisch
Beschreibung:

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 andFunction Characterization: Methods and Protocols provide different 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.
Includes cutting-edge methods and protocols
Recombinant Overexpression of Mammalian TSPO Isoforms 1 and 2.- Functional Assembly of Soluble and Membrane Recombinant Proteins of Mammalian NADPH Oxidase Complex.- Direct Extraction and Purification of Recombinant Membrane Proteins from Pichia pastoris protoplasts.- Cell-free Expression for the Study of Hydrophobic Proteins: The Example of Yeast ATP-synthase subunits.- Wheat Germ Cell Free Overexpression for the Production of Membrane Proteins.- Methyl-specific Isotope Labelling Strategies for NMR Studies of Membrane Proteins.- Labeling of Membrane Complexes for Electron Microscopy.- Expression, Biochemistry, and Stabilization with Camel Antibodies of Membrane Proteins Case Study of The Mouse 5-HT Receptor.- Characterization of New Detergent and Detergent Mimetics by Scattering Techniques for Membrane Protein Crystallization.- Secondary Structure Determination by Means of ATR-FTIR Spectroscopy.- Native Mass Spectroscopy forthe Characterization of Structure and Interactions of Membrane Proteins.- Mass Spectroscopy of Mitochondrial Membrane Proteins Complexes.- 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.- Reconstitution of Membrane Protein in Liposomes.- Ion Channels as Reporters of Membrane Receptor Function: Automated Analysis in Xenopus Oocytes.- The CRACAM Robot: Two Dimensional Crystallization of Membrane Protein.- Reconstitution of Membrane Proteins into Nanodiscs for Single Particle Electron Microscopy.- Solid-state NMR of Membrane Protein Reconstituted in Proteoliposomes, the Case of TSPO.- Sample Preparation for Membrane Protein Structural Studies by Solid-State NMR.- Simulation of Ligand Binding To Membrane Proteins.- 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.

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