title: Nano-scale alignment of proteins on a flexible DNA backbone.
abstract: Nano-scale alignment of several proteins with freedom of motion is equivalent to an enormous increase in effective local concentration of proteins and will enable otherwise impossible weak and/or cooperative associations between them or with their ligands. For this purpose, a DNA backbone made of six oligodeoxynucleotide (ODN) chains is designed in which five double-stranded segments are connected by four single-stranded flexible linkers. A desired protein with an introduced cysteine is connected covalently to the 5'-end of azido-ODN by catalyst-free click chemistry. Then, six protein-ODN conjugates are assembled with their complementary nucleotide sequences into a single multi-protein-DNA complex, and six proteins are aligned along the DNA backbone. Flexible alignment of proteins is directly observed by high-speed AFM imaging, and association of proteins with weak interaction is demonstrated by fluorescence resonance energy transfer between aligned proteins.
authors.affiliation: Department of Molecular Biosciences, Kyoto Sangyo University, Kyoto, Japan. nojima.t.aa@m.titech.ac.jp
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Nojima
authors.forename: Hiroki
authors.initials: H
authors.lastname: Konno
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kodera
authors.forename: Kohji
authors.initials: K
authors.lastname: Seio
authors.forename: Hideki
authors.initials: H
authors.lastname: Taguchi
authors.forename: Masasuke
authors.initials: M
authors.lastname: Yoshida
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism*; D018625:Microscopy, Atomic Force; D008958:Models, Molecular; D036103:Nanotechnology; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation
chemical_list: C485182:Cyan Fluorescent Protein; D009838:Oligodeoxyribonucleotides; D049452:Green Fluorescent Proteins; D004247:DNA