title: Patterning protein complexes on DNA nanostructures using a GFP nanobody.
abstract: DNA nanostructures have become an important and powerful tool for studying protein function over the last 5 years. One of the challenges, though, has been the development of universal methods for patterning protein complexes on DNA nanostructures. Herein, we present a new approach for labeling DNA nanostructures by functionalizing them with a GFP nanobody. We demonstrate the ability to precisely control protein attachment via our nanobody linker using two enzymatic model systems, namely adenylyl cyclase activity and myosin motility. Finally, we test the power of this attachment method by patterning unpurified, endogenously expressed Arp2/3 protein complex from cell lysate. By bridging DNA nanostructures with a fluorescent protein ubiquitous throughout cell and developmental biology and protein biochemistry, this approach significantly streamlines the application of DNA nanostructures as a programmable scaffold in biological studies.
authors.affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, Minnesota, 55455.
authors.forename: R F
authors.initials: RF
authors.lastname: Sommese
authors.affiliation: Department of Physics, Arizona State University, Tempe, Arizona, 85287.|Biodesign Center for Molecular Design and Biomimetics, Arizona State University, Tempe, Arizona, 85287.
authors.forename: R F
authors.initials: RF
authors.lastname: Hariadi
authors.affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, Minnesota, 55455.
authors.forename: K
authors.initials: K
authors.lastname: Kim
authors.affiliation: Biodesign Center for Molecular Design and Biomimetics, Arizona State University, Tempe, Arizona, 85287.|School of Molecular Sciences, Arizona State University, Tempe, Arizona, 85287.
authors.forename: M
authors.initials: M
authors.lastname: Liu
authors.affiliation: Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, Tennessee, 37232.
authors.forename: M J
authors.initials: MJ
authors.lastname: Tyska
authors.affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, Minnesota, 55455. sivaraj@umn.edu.
authors.forename: S
authors.initials: S
authors.lastname: Sivaramakrishnan
mesh_terms: D051376:Actin-Related Protein 2-3 Complex / Q000737:chemistry*; D000818:Animals; D004247:DNA / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D061905:Single-Domain Antibodies / Q000737:chemistry*; D013552:Swine
chemical_list: D051376:Actin-Related Protein 2-3 Complex; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies; D049452:Green Fluorescent Proteins; D004247:DNA
keywords: Arp2/3; DNA nanostructures; GFP; adenylyl cyclase; myosin VI; nanobody