Patterning protein complexes on DNA nanostructures using a GFP nanobody

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 n...

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Bibliographic Details
Published in:Protein science 2016-11, Vol.25 (11), p.2089-2094
Main Authors: Sommese, R. F., Hariadi, R. F., Kim, K., Liu, M., Tyska, M. J., Sivaramakrishnan, S.
Format: Article
Language:English
Subjects:
Actin-Related Protein 2-3 Complex - chemistry
adenylyl cyclase
Animals
Arp2/3
Arp2/3 protein
Attachment
Biochemistry
Biology
Bridging
Control systems
Deoxyribonucleic acid
Developmental biology
DNA
DNA - chemistry
DNA nanostructures
Fluorescence
GFP
Green fluorescent protein
Green Fluorescent Proteins - chemistry
Labels
Marking
Methods and Applications
Myosin
myosin VI
Nanobodies
nanobody
Nanostructure
Nanostructures - chemistry
Pattern formation
Patterning
Proteins
Recombinant Fusion Proteins - chemistry
Single-Domain Antibodies - chemistry
Stability
Swine
Test procedures
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Summary: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.
ISSN:0961-8368
1469-896X
DOI:10.1002/pro.3020