title: Self-sorting of foreign proteins in a bacterial nanocompartment.
abstract: Nature uses bottom-up approaches for the controlled assembly of highly ordered hierarchical structures with defined functionality, such as organelles, molecular motors, and transmembrane pumps. The field of bionanotechnology draws inspiration from nature by utilizing biomolecular building blocks such as DNA, proteins, and lipids, for the (self-) assembly of new structures for applications in biomedicine, optics, or electronics. Among the toolbox of available building blocks, proteins that form cage-like structures, such as viruses and virus-like particles, have been of particular interest since they form highly symmetrical assemblies and can be readily modified genetically or chemically both on the outer or inner surface. Bacterial encapsulins are a class of nonviral protein cages that self-assemble in vivo into stable icosahedral structures. Using teal fluorescent proteins (TFP) engineered with a specific native C-terminal docking sequence, we report the molecular self-sorting and selective packaging of TFP cargo into bacterial encapsulins during in vivo assembly. Using native mass spectrometry techniques, we show that loading of either monomeric or dimeric TFP cargo occurs with unprecedented high fidelity and exceptional loading accuracy. Such self-assembling systems equipped with self-sorting capabilities would open up exciting opportunities in nanotechnology, for example, as artificial (molecular storage or detoxification) organelles or as artificial cell factories for in situ biocatalysis.
authors: Laboratory for Biomolecular Nanotechnology, MESA+ Institute for Nanotechnology, University of Twente , P.O. Box 217, 7500 AE Enschede, The Netherlands.
authors: W Frederik
authors: WF
authors: Rurup
authors: Joost
authors: J
authors: Snijder
authors: Melissa S T
authors: MS
authors: Koay
authors: Albert J R
authors: AJ
authors: Heck
authors: Jeroen J L M
authors: JJ
authors: Cornelissen
mesh_terms: D001951:Brevibacterium / Q000166:cytology* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000378:metabolism*; D008958:Models, Molecular; D055503:Protein Multimerization; D021381:Protein Transport; D020124:Thermotoga maritima / Q000166:cytology* / Q000378:metabolism
chemical_list: C485182:Cyan Fluorescent Protein; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins