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Angew Chem Int Ed Engl. 2016 Mar 18;55(13):4348-52. doi: 10.1002/anie.201510313. Epub 2016 Feb 24.

Orthogonal Protein Assembly on DNA Nanostructures Using Relaxases.

Author information

1
Departamento de Biología Molecular e Instituto de Biomedicina y Biotecnología de Cantabria, Universidad de Cantabria-Consejo Superior de Investigaciones Científicas-SODERCAN, Albert Einstein 22, 39011, Santander, Spain.
2
Physik-Department E14 and ZNN/WSI, TU Munich, Am Coulombwall 4a, 85748, Garching, Germany.
3
Physik-Department E14 and ZNN/WSI, TU Munich, Am Coulombwall 4a, 85748, Garching, Germany. simmel@tum.de.
4
Departamento de Biología Molecular e Instituto de Biomedicina y Biotecnología de Cantabria, Universidad de Cantabria-Consejo Superior de Investigaciones Científicas-SODERCAN, Albert Einstein 22, 39011, Santander, Spain. delacruz@unican.es.

Abstract

DNA-binding proteins are promising reagents for the sequence-specific modification of DNA-based nanostructures. Here, we investigate the utility of a series of relaxase proteins-TrwC, TraI, and MobA-for nanofunctionalization. Relaxases are involved in the conjugative transfer of plasmids between bacteria, and bind to their DNA target sites via a covalent phosphotyrosine linkage. We study the binding of the relaxases to two standard DNA origami structures-rodlike six-helix bundles and flat rectangular origami sheets. We find highly orthogonal binding of the proteins with binding yields of 40-50 % per binding site, which is comparable to other functionalization methods. The yields differ for the two origami structures and also depend on the position of the binding sites. Due to their specificity for a single-stranded DNA target, their orthogonality, and their binding properties, relaxases are a uniquely useful addition to the toolbox available for the modification of DNA nanostructures with proteins.

KEYWORDS:

DNA nanotechnology; DNA origami; bacterial conjugation; bioconjugation; relaxase

PMID:
26915475
PMCID:
PMC5067690
DOI:
10.1002/anie.201510313
[Indexed for MEDLINE]
Free PMC Article

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