title: Modified TMV Particles as Beneficial Scaffolds to Present Sensor Enzymes.
abstract: Tobacco mosaic virus (TMV) is a robust nanotubular nucleoprotein scaffold increasingly employed for the high density presentation of functional molecules such as peptides, fluorescent dyes, and antibodies. We report on its use as advantageous carrier for sensor enzymes. A TMV mutant with a cysteine residue exposed on every coat protein (CP) subunit (TMVCys) enabled the coupling of bifunctional maleimide-polyethylene glycol (PEG)-biotin linkers (TMVCys/Bio). Its surface was equipped with two streptavidin [SA]-conjugated enzymes: glucose oxidase ([SA]-GOx) and horseradish peroxidase ([SA]-HRP). At least 50% of the CPs were decorated with a linker molecule, and all thereof with active enzymes. Upon use as adapter scaffolds in conventional "high-binding" microtiter plates, TMV sticks allowed the immobilization of up to 45-fold higher catalytic activities than control samples with the same input of enzymes. Moreover, they increased storage stability and reusability in relation to enzymes applied directly to microtiter plate wells. The functionalized TMV adsorbed to solid supports showed a homogeneous distribution of the conjugated enzymes and structural integrity of the nanorods upon transmission electron and atomic force microscopy. The high surface-increase and steric accessibility of the viral scaffolds in combination with the biochemical environment provided by the plant viral coat may explain the beneficial effects. TMV can, thus, serve as a favorable multivalent nanoscale platform for the ordered presentation of bioactive proteins.
authors.affiliation: Department of Molecular Biology and Plant Virology, Institute of Biomaterials and Biomolecular Systems, University of Stuttgart Stuttgart, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Koch
authors.affiliation: Department of Molecular Biology and Plant Virology, Institute of Biomaterials and Biomolecular Systems, University of Stuttgart Stuttgart, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Wabbel
authors.affiliation: Department of Molecular Biology and Plant Virology, Institute of Biomaterials and Biomolecular Systems, University of Stuttgart Stuttgart, Germany.
authors.forename: Fabian J
authors.initials: FJ
authors.lastname: Eber
authors.affiliation: Chemistry of Oxydic and Organic Interfaces, Karlsruhe Institute of Technology, Institute of Functional Interfaces Karlsruhe, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Krolla-Sidenstein
authors.affiliation: Chemistry of Oxydic and Organic Interfaces, Karlsruhe Institute of Technology, Institute of Functional Interfaces Karlsruhe, Germany.
authors.forename: Carlos
authors.initials: C
authors.lastname: Azucena
authors.affiliation: Chemistry of Oxydic and Organic Interfaces, Karlsruhe Institute of Technology, Institute of Functional Interfaces Karlsruhe, Germany.
authors.forename: Hartmut
authors.initials: H
authors.lastname: Gliemann
authors.affiliation: Department of Molecular Biology and Plant Virology, Institute of Biomaterials and Biomolecular Systems, University of Stuttgart Stuttgart, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Eiben
authors.affiliation: Department of New Materials and Biosystems, Max-Planck-Institute for Intelligent Systems Stuttgart, Germany.
authors.forename: Fania
authors.initials: F
authors.lastname: Geiger
authors.affiliation: Department of Molecular Biology and Plant Virology, Institute of Biomaterials and Biomolecular Systems, University of Stuttgart Stuttgart, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Wege
keywords: biotemplate; enzyme cascade system; enzyme shelf-life; glucose sensor; reusability; scaffold; surface immobilization; tobacco mosaic virus (TMV)