- Sommese RF, Hariadi RF, Kim K, Liu M, Tyska MJ, Sivaramakrishnan S. Patterning Protein Complexes on DNA Nanostructures Using a GFP-Nanobody. Protein science : a publication of the Protein Society. 2016 Aug 18. PMID: 27538185 [PubMed].
DNA nanostructures have become an important and powerful tool for studying protein function over the last five 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 nanostructure as a programmable scaffold in biological studies. This article is protected by copyright. All rights reserved.