Titlebook: Expression, Purification, and Structural Biology of Membrane Proteins; Camilo Perez,Timm Maier Book 2020 Springer Science+Business Media,

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Expression and Purification of Membrane Proteins in ,tochondrial membrane proteins developed in our laboratory during the last 15?years. To optimize their expression in a functional form, different promoter systems as well as codon-optimization and complementation strategies were established. Purification approaches were developed which remove the mem

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Membrane Protein Expression in Insect Cells Using the Baculovirus Expression Vector System,st. Recombinant protein production is the first step in the protein tool generation for biochemical and biophysical studies. Here, we provide simplified protocols that facilitate the generation of high-quality virus and initial expression analysis for integral membrane protein targets utilizing the

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Membrane Protein Solubilization and Quality Control: An Example of a Primary Active Transporter,o identify the best-suited detergent, which on the one hand would solubilize large amounts of the target protein but on the other hand would sustain the protein’s activity. Here we describe the solubilization screen and subsequent activity assay we have optimized for the bacterial P-type ATPase KdpF

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GPCR Solubilization and Quality Control,ions, making them interesting pharmacological targets. In order to study their structure and function, GPCRs are traditionally extracted from membranes using detergents. However, due to their hydrophobic nature, intrinsic instability in aqueous solutions, and their?denaturing effects, the?isolation

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Affinity Purification of Membrane Proteins,s. One of the most powerful methods for isolation of the membrane protein of interest is affinity purification. This methodology typically relies on engineering an affinity tag into the protein of interest and an affinity resin that specifically recognizes the tag, allowing one to purify the target

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Purification of Membrane Proteins by Affinity Chromatography with On-Column Protease Cleavage,t membrane protein is cloned into an expression plasmid and then introduced into . cells for overexpression. Membranes from bacterial cells are isolated and the tagged target membrane protein is solubilized in detergent and subsequently bound to an affinity matrix. Tagged proteins are commonly elute