Sagredo Sandra, Pirzer Tobias, Aghebat Rafat Ali, Goetzfried Marisa A, Moncalian Gabriel, Simmel Friedrich C, de la Cruz Fernando
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.
Physik-Department E14 and ZNN/WSI, TU Munich, Am Coulombwall 4a, 85748, Garching, Germany.
Angew Chem Int Ed Engl. 2016 Mar 18;55(13):4348-52. doi: 10.1002/anie.201510313. Epub 2016 Feb 24.
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.
DNA结合蛋白是用于对基于DNA的纳米结构进行序列特异性修饰的有前景的试剂。在此，我们研究了一系列松弛酶蛋白——TrwC、TraI和MobA——用于纳米功能化的效用。松弛酶参与细菌间质粒的接合转移，并通过共价磷酸酪氨酸连接与它们的DNA靶位点结合。我们研究了松弛酶与两种标准DNA折纸结构——棒状六螺旋束和平坦矩形折纸片——的结合。我们发现这些蛋白具有高度正交的结合，每个结合位点的结合产率为40 - 50%，这与其他功能化方法相当。两种折纸结构的产率不同，并且还取决于结合位点的位置。由于它们对单链DNA靶标的特异性、正交性以及结合特性，松弛酶是蛋白质修饰DNA纳米结构可用工具库中独特且有用的补充。
