Patterning protein complexes on DNA nanostructures using a GFP nanobody.

Sommese RF, Hariadi RF, Kim K, Liu M, Tyska MJ, Sivaramakrishnan S
Protein Sci. 2016 25 (11): 2089-2094

PMID: 27538185 · PMCID: PMC5079250 · DOI:10.1002/pro.3020

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.

MeSH Terms (8)

Actin-Related Protein 2-3 Complex Animals DNA Green Fluorescent Proteins Nanostructures Recombinant Fusion Proteins Single-Domain Antibodies Swine

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