Albany 2013: Book of Abstracts
June 11-15 2013
©Adenine Press (2012)
Arranging enzymes and receptors with nanometer-scale precision on molecular switchboards
DNA is a useful material for constructing nanoscale structures in nearly any three-dimensional shape desired. The DNA nanostructure can also be equipped with specific docking sites for proteins. Cellular processes and chemical transformations take place in several reaction steps. Multiple enzymes cooperate in specific fashion to catalyze the sequential chemical transformation steps. Such natural systems are effectively reconstructed in vitro if the individual enzymes locate in the correct relative orientations. DNA-origami structures can be used as “molecular switchboards” to arrange enzymes and other proteins with nanometer-scale precision.
A new method was developed for locating the proteins by means of special “adapters” known as zinc-finger proteins based only on proteins. Zinc fingers are suitable site-selective adapters for targeting specific locations within DNA-origami structures. Several different adapters carrying different proteins can independently bind at defined locations on this type of nanostructure. A basic leucine zipper (bZIP) protein is also a candidate for the site-selective adaptor. A well-characterized bZIP protein GCN4 was chosen as an adaptor for specific addresses. Analyses by atomic force microscopy and gel electrophoreses demonstrate specific binding of GCN4 adaptor to the addresses containing the GCN4 binding sites on DNA origami.
The adaptor derived from GCN4 and that form a zinc finger protein zif268, for which we have reported previously, acted as orthogonal adaptors to the respective addresses on DNA origami. Therefore these orthogonal adaptors would be useful to place multiple engineered proteins at different addresses on DNA origami. Especially, the homodimeric nature of GCN4 adaptor is indispensable for constructing the assembly of the naturally abundant dimeric proteins and/or enzymes to efficiently carry out chemical reactions and signal transductions in vitro on DNA origami.
Nakata, E.; Liew, F. F.; Uwatoko, C. Kiyonaka, S. Mori, Y.; Katsuda, Y.; Endo, M.; Sugiyama, H.; Morii T. (2012) Zinc finger proteins for site-specific protein positioning on DNA origami. Angew Chem. Int. Ed. 51, 2421–2424.
Ngo Anh Tien
Institute of Advanced Energy