title: Protein-DNA chimeras for nano assembly.
abstract: In synthetic biology, "understanding by building" requires exquisite control of the molecular constituents and their spatial organization. Site-specific coupling of DNA to proteins allows arrangement of different protein functionalities with emergent properties by self-assembly on origami-like DNA scaffolds or by direct assembly via Single-Molecule Cut & Paste (SMC&P). Here, we employed the ybbR-tag/Sfp system to covalently attach Coenzyme A-modified DNA to GFP and, as a proof of principle, arranged the chimera in different patterns by SMC&P. Fluorescence recordings of individual molecules proved that the proteins remained folded and fully functional throughout the assembly process. The high coupling efficiency and specificity as well as the negligible size (11 amino acids) of the ybbR-tag represent a mild, yet versatile, general and robust way of adding a freely programmable and highly selective attachment site to virtually any protein of interest.
authors.affiliation: Center for Nanoscience and Department of Physics, University of Munich , Amalienstraße 54, 80799 Munich, Germany.
authors.forename: Diana A
authors.initials: DA
authors.lastname: Pippig
authors.forename: Fabian
authors.initials: F
authors.lastname: Baumann
authors.forename: Mathias
authors.initials: M
authors.lastname: Strackharn
authors.forename: Daniela
authors.initials: D
authors.lastname: Aschenbrenner
authors.forename: Hermann E
authors.initials: HE
authors.lastname: Gaub
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D003065:Coenzyme A / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D049329:Nanostructures / Q000737:chemistry*; D009690:Nucleic Acid Conformation; D013194:Staining and Labeling; D013499:Surface Properties; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D055767:Immobilized Proteins; C082662:phosphopantetheinyl transferase; D049452:Green Fluorescent Proteins; D004247:DNA; D017855:Transferases (Other Substituted Phosphate Groups); D003065:Coenzyme A