title: A simple tagging system for protein encapsulation.
abstract: Molecular containers that encapsulate specific cargo can be useful for many natural and non-natural processes. We report a simple system, based on charge complementarity, for the encapsulation of appropriately tagged proteins within an engineered, proteinaceous capsid. Four negative charges per monomer were added to the lumazine synthase from Aquifex aeolicus (AaLS). The capsids formed by the engineered AaLS associate with green fluorescent protein bearing a positively charged deca-arginine tag upon coproduction in Escherichia coli. Analytical ultracentrifugation and scanning force microscopy studies indicated that the engineered AaLS retains the ability to form capsids, but that their average size was substantially increased. The success of this strategy demonstrates that both the container and guest components of protein-based encapsulation systems can be convergently designed in a straightforward manner, which may help to extend their versatility.
authors.affiliation: Laboratorium für Organische Chemie, ETH Zürich, Hönggerberg HCI F337, CH-8093, Zürich, Switzerland.
authors.forename: Florian P
authors.initials: FP
authors.lastname: Seebeck
authors.forename: Kenneth J
authors.initials: KJ
authors.lastname: Woycechowsky
authors.forename: Wei
authors.initials: W
authors.lastname: Zhuang
authors.forename: Jürgen P
authors.initials: JP
authors.lastname: Rabe
authors.forename: Donald
authors.initials: D
authors.lastname: Hilvert
mesh_terms: D019943:Amino Acid Substitution; D001120:Arginine / Q000737:chemistry; D001419:Bacteria / Q000201:enzymology; D002850:Chromatography, Gel; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics; D015202:Protein Engineering
chemical_list: D009097:Multienzyme Complexes; D049452:Green Fluorescent Proteins; C044822:6,7-dimethyl-8-ribityllumazine synthase; D001120:Arginine