title: Protein immobilization on liposomes and lipid-coated nanoparticles by protein trans-splicing.
abstract: A plethora of methods exist to link proteins to surfaces in order to generate functionalized materials. However, general tools that lead to functional immobilization of recombinantly expressed proteins on membranes such as liposomes or lipid-coated nanoparticles are rare. Here we present an approach that takes advantage of a double-palmitoylated peptide that mediates stable membrane anchoring in combination with protein trans-splicing for efficient immobilization of recombinant proteins fused to split intein segments. Two different DnaE split inteins from Synechocystis and Nostoc punctiforme are tested and compared to immobilization via direct native chemical ligation using a protein thioester. Protein trans-splicing proceeds at low protein concentrations and leads to functionalized vesicles and membrane-coated silica nanoparticles.
authors.affiliation: Center for Integrated Protein Science Munich and Technische Universität München, Fakultät Chemie, Garching, Germany.
authors.forename: Nam Ky
authors.initials: NK
authors.lastname: Chu
authors.forename: Diana
authors.initials: D
authors.lastname: Olschewski
authors.forename: Ralf
authors.initials: R
authors.lastname: Seidel
authors.forename: Konstanze F
authors.initials: KF
authors.lastname: Winklhofer
authors.forename: Jörg
authors.initials: J
authors.lastname: Tatzelt
authors.forename: Martin
authors.initials: M
authors.lastname: Engelhard
authors.forename: Christian F W
authors.initials: CF
authors.lastname: Becker
mesh_terms: D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D047668:Inteins / Q000235:genetics; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000378:metabolism*; D008958:Models, Molecular; D053758:Nanoparticles / Q000737:chemistry*; D046937:Nostoc / Q000235:genetics; D019154:Protein Splicing; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D046939:Synechocystis / Q000235:genetics; D020040:Trans-Splicing
chemical_list: D055767:Immobilized Proteins; D008055:Lipids; D008081:Liposomes; D011993:Recombinant Fusion Proteins