title: General strategy for ordered noncovalent protein assembly on well-defined nanoscaffolds.
abstract: Here we develop a novel approach allowing the noncovalent assembly of proteins on well-defined nanoscaffolds such as virus particles. The antibody-binding peptide Z33 was genetically fused to the monomeric yellow fluorescent protein and 4-coumarate:CoA-ligase 2. This Z33 "tag" allowed their patterning on the surface of zucchini yellow mosaic virus by means of specific antibodies directed against the coat protein of the virus. The approach was validated by affinity assays and correlative microscopy. The coverage efficiency was ≈ 87%. Fluorescence and enzymatic activity were fully retained after assembly. The principle of using the combination of a scaffold-specific antibody and Z33-fusion proteins can be extended to a wide variety of proteins/enzymes and antigenic scaffolds to support coupling for creating functional "biochips" with optical or catalytic properties.
authors.affiliation: UMR 1332 Biologie du Fruit et Pathologie, INRA-Université Bordeaux 2 , 71, av. Edouard Bourlaux, CS 20032-33882 Villenave d'Ornon Cedex, France.
authors.forename: Jan
authors.initials: J
authors.lastname: Pille
authors.forename: Daniela
authors.initials: D
authors.lastname: Cardinale
authors.forename: Noëlle
authors.initials: N
authors.lastname: Carette
authors.forename: Carmelo
authors.initials: C
authors.lastname: Di Primo
authors.forename: Jane
authors.initials: J
authors.lastname: Besong-Ndika
authors.forename: Jocelyne
authors.initials: J
authors.lastname: Walter
authors.forename: Hervé
authors.initials: H
authors.lastname: Lecoq
authors.forename: Mark B
authors.initials: MB
authors.lastname: van Eldijk
authors.forename: Ferdinanda C M
authors.initials: FC
authors.lastname: Smits
authors.forename: Sanne
authors.initials: S
authors.lastname: Schoffelen
authors.forename: Jan C M
authors.initials: JC
authors.lastname: van Hest
authors.forename: Kristiina
authors.initials: K
authors.lastname: Mäkinen
authors.forename: Thierry
authors.initials: T
authors.lastname: Michon
mesh_terms: D029681:Arabidopsis Proteins / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry; D036022:Capsid Proteins / Q000737:chemistry*; D003066:Coenzyme A Ligases / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry; D007700:Kinetics; D008164:Luminescent Proteins / Q000737:chemistry; D046529:Microscopy, Electron, Transmission; D009029:Mosaic Viruses / Q000737:chemistry; D049329:Nanostructures / Q000737:chemistry*; D010316:Particle Size; D015202:Protein Engineering; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D014771:Virion / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D029681:Arabidopsis Proteins; D001426:Bacterial Proteins; D036022:Capsid Proteins; D004800:Enzymes, Immobilized; D007074:Immunoglobulin G; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D003066:Coenzyme A Ligases