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Membrane protein synthesis in cell-free systems: From bio-mimetic systems to bio-membranes
Authors:Rita Sachse  Srujan K. Dondapati  Susanne F. Fenz  Thomas Schmidt  Stefan Kubick
Affiliation:1. Fraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses Potsdam-Golm, Am Mühlenberg 13, 14476 Potsdam, Germany;2. Department of Cell and Developmental Biology, Biocenter, University of Würzburg, Am Hubland, 97074 Würzburg, Germany;3. Leiden Institute of Physics, Leiden University, PO Box 9504, 2300 RA Leiden, The Netherlands
Abstract:When taking up the gauntlet of studying membrane protein functionality, scientists are provided with a plethora of advantages, which can be exploited for the synthesis of these difficult-to-express proteins by utilizing cell-free protein synthesis systems. Due to their hydrophobicity, membrane proteins have exceptional demands regarding their environment to ensure correct functionality. Thus, the challenge is to find the appropriate hydrophobic support that facilitates proper membrane protein folding. So far, various modes of membrane protein synthesis have been presented. Here, we summarize current state-of-the-art methodologies of membrane protein synthesis in biomimetic-supported systems. The correct folding and functionality of membrane proteins depend in many cases on their integration into a lipid bilayer and subsequent posttranslational modification. We highlight cell-free systems utilizing the advantages of biological membranes.
Keywords:a, membrane anchored   c, multimeric protein complex   CF, cell-free   CMC, critical micelle concentration   EF, elongation factor   ER, endoplasmic reticulum   GUVs, giant unilamellar vesicles   IF, initiation factor   LUVs, large unilamellar vesicles   MP, membrane protein   n.d., not determined   PURE system, protein synthesis using recombinant elements system   Ref., reference   RF, release factor   RRL, rabbit reticulocyte lysate   Sf, Spodoptera frugiperda   TMR, transmembrane region   WG, wheat germ
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