Patterning protein complexes on DNA nanostructures using a GFP nanobody.

Autor: Sommese RF; Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, Minnesota, 55455., Hariadi RF; Department of Physics, Arizona State University, Tempe, Arizona, 85287.; Biodesign Center for Molecular Design and Biomimetics, Arizona State University, Tempe, Arizona, 85287., Kim K; Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, Minnesota, 55455., Liu M; Biodesign Center for Molecular Design and Biomimetics, Arizona State University, Tempe, Arizona, 85287.; School of Molecular Sciences, Arizona State University, Tempe, Arizona, 85287., Tyska MJ; Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, Tennessee, 37232., Sivaramakrishnan S; Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, Minnesota, 55455. sivaraj@umn.edu.
Jazyk: angličtina
Zdroj: Protein science : a publication of the Protein Society [Protein Sci] 2016 Nov; Vol. 25 (11), pp. 2089-2094. Date of Electronic Publication: 2016 Aug 31.
DOI: 10.1002/pro.3020
Abstrakt: 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.
(© 2016 The Protein Society.)
Databáze: MEDLINE
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