title: Inspired and stabilized by nature: ribosomal synthesis of the human voltage gated ion channel (VDAC) into 2D-protein-tethered lipid interfaces.
abstract: We present an elegant synthesis and reconstitution approach for functional studies of voltage responsive membrane proteins. For such studies, we propose a planar architecture of an S-layer-supported lipid membrane as a suitable matrix for presenting unmodified membrane protein species, and here we focus on the voltage-dependent anion channel (VDAC) from human mitochondria. The presented cell-free strategy, in which VDAC proteins are synthesized in bacterial cell lysate, into a membrane structure, offers a great advantage in the study of such subtle membrane proteins over the conventional, cell-based synthesis approach in terms of reproducibility. The material-assay combination is superior over cell-culture related synthesis and purification approaches as here we bypass by a one-step synthesis procedure the complex cell culture, and expression and purification endeavours, and, moreover, the protein of interest never has to be detergent solubilized and had been synthesized de novo. We provide here a detailed description from the all over procedure and our first results, describing in detail the cell-free synthesis and robustness of such a material-assay combination: functional VDAC protein species embedded in a planar membrane architecture and ready for electrochemical characterization.
authors.affiliation: Institute for Synthetic Bioarchitectures, Department of NanoBiotechnology, University of Natural Resources and Life Sciences, Muthgasse 11, Vienna 1190, Austria. eva.sinner@boku.ac.at.
authors.forename: Samar
authors.initials: S
authors.lastname: Damiati
authors.forename: Sonja
authors.initials: S
authors.lastname: Zayni
authors.forename: Angelika
authors.initials: A
authors.lastname: Schrems
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kiene
authors.forename: Uwe B
authors.initials: UB
authors.lastname: Sleytr
authors.forename: Joël
authors.initials: J
authors.lastname: Chopineau
authors.forename: Bernhard
authors.initials: B
authors.lastname: Schuster
authors.forename: Eva-Kathrin
authors.initials: EK
authors.lastname: Sinner
mesh_terms: D055664:Electrochemical Techniques; D006801:Humans; D007094:Imides / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D008055:Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008928:Mitochondria / Q000737:chemistry* / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry; D011437:Propylamines / Q000737:chemistry; D012270:Ribosomes / Q000737:chemistry*; D050994:Voltage-Dependent Anion Channels / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D007094:Imides; D008051:Lipid Bilayers; D008055:Lipids; D008565:Membrane Proteins; D010714:Phosphatidylethanolamines; D011437:Propylamines; D050994:Voltage-Dependent Anion Channels; C000613388:1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride; C018525:1,2-dimyristoylphosphatidylethanolamine