title: Designer amphiphilic proteins as building blocks for the intracellular formation of organelle-like compartments.
abstract: Nanoscale biological materials formed by the assembly of defined block-domain proteins control the formation of cellular compartments such as organelles. Here, we introduce an approach to intentionally 'program' the de novo synthesis and self-assembly of genetically encoded amphiphilic proteins to form cellular compartments, or organelles, in Escherichia coli. These proteins serve as building blocks for the formation of artificial compartments in vivo in a similar way to lipid-based organelles. We investigated the formation of these organelles using epifluorescence microscopy, total internal reflection fluorescence microscopy and transmission electron microscopy. The in vivo modification of these protein-based de novo organelles, by means of site-specific incorporation of unnatural amino acids, allows the introduction of artificial chemical functionalities. Co-localization of membrane proteins results in the formation of functionalized artificial organelles combining artificial and natural cellular function. Adding these protein structures to the cellular machinery may have consequences in nanobiotechnology, synthetic biology and materials science, including the constitution of artificial cells and bio-based metamaterials.
authors.affiliation: 1] Institute for Macromolecular Chemistry, University of Freiburg, Stefan-Meier-Str. 31 D-79104 Freiburg, Germany [2] Institute for Pharmaceutical Sciences, University of Freiburg, Albertstr. 25 D-79104 Freiburg, Germany [3] Freiburg Institute for Advanced Studies (FRIAS), School of Soft Matter Research, University of Freiburg, Albertstr. 19 D-79104 Freiburg, Germany [4] Faculty of Chemistry and Pharmacy, University of Freiburg, Fahnenbergplatz D-79104 Freiburg, Germany.
authors.forename: Matthias C
authors.initials: MC
authors.lastname: Huber
authors.affiliation: 1] Institute for Macromolecular Chemistry, University of Freiburg, Stefan-Meier-Str. 31 D-79104 Freiburg, Germany [2] Institute for Pharmaceutical Sciences, University of Freiburg, Albertstr. 25 D-79104 Freiburg, Germany [3] Freiburg Institute for Advanced Studies (FRIAS), School of Soft Matter Research, University of Freiburg, Albertstr. 19 D-79104 Freiburg, Germany [4] Faculty of Biology, University of Freiburg, Schänzlestrasse 1 D-79085 Freiburg, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Schreiber
authors.affiliation: 1] Bio- and Nano-Photonics, Department of Microsystems Engineering, University of Freiburg, Georges-Köhler-Allee 102 D-79110 Freiburg, Germany [2] BIOSS Centre for Biological Signalling Studies, University of Freiburg, Schänzlestrasse 18 D-79104 Freiburg, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: von Olshausen
authors.affiliation: Chemical Biology Research Group, Hungarian Academy of Sciences, CNS, IOC, Magyar tudósok krt. 2 H-1117 Budapest, Hungary.
authors.forename: Balázs R
authors.initials: BR
authors.lastname: Varga
authors.affiliation: BIOSS Centre for Biological Signalling Studies, University of Freiburg, Schänzlestrasse 18 D-79104 Freiburg, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Kretz
authors.affiliation: Institute for Neuroanatomy University of Freiburg, Albertstr. 17 D-79104 Freiburg, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Joch
authors.affiliation: 1] Faculty of Chemistry and Pharmacy, University of Freiburg, Fahnenbergplatz D-79104 Freiburg, Germany [2] Institute of Pharmaceutical Sciences, Department of Pharmaceutical Technology and Biopharmacy, University of Freiburg, Hermann-Herder-Str. 9 D-79104 Freiburg, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Barnert
authors.affiliation: 1] Faculty of Chemistry and Pharmacy, University of Freiburg, Fahnenbergplatz D-79104 Freiburg, Germany [2] BIOSS Centre for Biological Signalling Studies, University of Freiburg, Schänzlestrasse 18 D-79104 Freiburg, Germany [3] Institute of Pharmaceutical Sciences, Department of Pharmaceutical Technology and Biopharmacy, University of Freiburg, Hermann-Herder-Str. 9 D-79104 Freiburg, Germany.
authors.forename: Rolf
authors.initials: R
authors.lastname: Schubert
authors.affiliation: BIOSS Centre for Biological Signalling Studies, University of Freiburg, Schänzlestrasse 18 D-79104 Freiburg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Eimer
authors.affiliation: Chemical Biology Research Group, Hungarian Academy of Sciences, CNS, IOC, Magyar tudósok krt. 2 H-1117 Budapest, Hungary.
authors.forename: Péter
authors.initials: P
authors.lastname: Kele
authors.affiliation: 1] Institute for Macromolecular Chemistry, University of Freiburg, Stefan-Meier-Str. 31 D-79104 Freiburg, Germany [2] Institute for Pharmaceutical Sciences, University of Freiburg, Albertstr. 25 D-79104 Freiburg, Germany [3] Freiburg Institute for Advanced Studies (FRIAS), School of Soft Matter Research, University of Freiburg, Albertstr. 19 D-79104 Freiburg, Germany [4] Faculty of Chemistry and Pharmacy, University of Freiburg, Fahnenbergplatz D-79104 Freiburg, Germany [5] Faculty of Biology, University of Freiburg, Schänzlestrasse 1 D-79085 Freiburg, Germany [6] BIOSS Centre for Biological Signalling Studies, University of Freiburg, Schänzlestrasse 18 D-79104 Freiburg, Germany [7] IMTEK Department of Microsystems Engineering, University of Freiburg, Georges-Köhler-Allee 103 D-79110 Freiburg, Germany [8] Center for Biosystems Analysis (ZBSA), University of Freiburg, Habsburger Str. 49 D-79104 Freiburg, Germany.
authors.forename: Stefan M
authors.initials: SM
authors.lastname: Schiller
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D015388:Organelles / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins