Patterning protein complexes on DNA nanostructures using a GFP nanobody

R. F. Sommese, Rizal Hariadi, K. Kim, M. Liu, M. J. Tyska, S. Sivaramakrishnan

Research output: Contribution to journalArticle

2 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)2089-2094
Number of pages6
JournalProtein Science
Volume25
Issue number11
DOIs
StatePublished - Nov 1 2016

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Keywords

  • adenylyl cyclase
  • Arp2/3
  • DNA nanostructures
  • GFP
  • myosin VI
  • nanobody

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Sommese, R. F., Hariadi, R., Kim, K., Liu, M., Tyska, M. J., & Sivaramakrishnan, S. (2016). Patterning protein complexes on DNA nanostructures using a GFP nanobody. Protein Science, 25(11), 2089-2094. https://doi.org/10.1002/pro.3020