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Orthogonal Protein Assembly on DNA Nanostructures Using Relaxases
Author(s) -
Sagredo Sandra,
Pirzer Tobias,
Aghebat Rafat Ali,
Goetzfried Marisa A.,
Moncalian Gabriel,
Simmel Friedrich C.,
de la Cruz Fernando
Publication year - 2016
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201510313
Subject(s) - dna , dna origami , chemistry , covalent bond , binding site , dna nanotechnology , biophysics , dna binding protein , nanotechnology , combinatorial chemistry , biochemistry , biology , materials science , gene , organic chemistry , transcription factor
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.