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Preparation of Recombinant Membrane Proteins from Pichia pastoris for Molecular Investigations

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Guyot, Lucile | Hartmann, Lucie | Mohammed‐bouteben, Sarah | Caro, Lydia | Wagner, Renaud

Edité par CCSD ; Wiley -

Pichia pastoris is a eukaryotic microorganism reputed for its ability to mass‐produce recombinant proteins, including integral membrane proteins, for various applications. This article details a series of protocols that progress towards the production of integral membrane proteins, their extraction and purification in the presence of detergents, and their eventual reconstitution in lipid nanoparticles. These basic procedures can be further optimized to provide integral membrane protein samples that are compatible with a number of structural and/or functional investigations at the molecular level. Each protocol provides general guidelines, technical hints, and specific recommendations, and is illustrated with case studies corresponding to several representative mammalian proteins.

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