This second edition provides new and updated protocols on membrane protein biology. Chapters explore methods for cloning and expression of membrane proteins, membrane protein complexes in prokaryotic and eukaryotic systems, approaches for protein purification, nanobody applications, as well as biophysical characterization and much more. Written for the highly successful Methods in Molecular Biology series, 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 thorough, Expression, Purification, and Structure Biology of Membrane Proteins, Second Edition serves to guide and encourage young researchers and newcomers to the field to tackle bold new studies on membrane proteins. Chapter 11 is available open access under a CC-BY 4.0 license via link.springer.com.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt

Expression of Eukaryotic Membrane Proteins in Insect Cells .- Integrated Modular Workflows for Membrane Protein Production in HEK293 Cells.- Preparation of Membrane Extracts from Plant Tissues.- Detection and Localization of Type II Membrane Proteins by Endogenous Gene Tagging and Confocal Microscopy.- Size Exclusion Chromatography with Multi-Angle Light Scattering for Detergent-Solubilised Integral Membrane Proteins.- GFP-nanobody-based fluorescence-detection size-exclusion chromatography (FSEC) method for the expression screening of membrane and secreted proteins.- Mass photometry analysis of membrane proteins in detergent micelles.- HS-AFM Applications to Study Membrane Proteins.- Expression and Purification of Integral Membrane Enzymes for Functional and Structural Studies.- Expression, Purification, and Structural Characterization of Membrane Proteins: A case study of the Pseudomonas aeruginosa ABC Transporter MsbA.- Design and optimization of BRIL fusion constructs for membrane protein cryo-EM studies.- Structural Analysis of Membrane Proteins in Cell-Derived Microvesicles.- Sub-tomogram averaging of membrane complexes in organelle-derived vesicles.- Studying the protein-membrane interaction of a mycobacterial ABC transporter through coarse-grained simulations.- Assay to measure scrambling of dolichol-linked oligosaccharide in reconstituted vesicles.- Preparation of plasma membrane vesicles, lysosomes, proteoliposomes and alternative sample types for SSM-based Electrophysiology.