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A Rationally Designed Connector for Assembly of Protein‐Functionalized DNA Nanostructures
Author(s) -
Koßmann Katja J.,
Ziegler Cornelia,
Angelin Alessandro,
Meyer Rebecca,
Skoupi Marc,
Rabe Kersten S.,
Niemeyer Christof M.
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201600039
Subject(s) - dna , nanotechnology , dna nanotechnology , dna origami , nanostructure , self assembly , synthetic biology , chemistry , combinatorial chemistry , computational biology , materials science , biology , biochemistry
We report on the rational engineering of the binding interface of the self‐ligating HaloTag protein to generate an optimized linker for DNA nanostructures. Five amino acids positioned around the active‐site entry channel for the chlorohexyl ligand (CH) of the HaloTag protein were exchanged for positively charged lysine amino acids to produce the HOB (halo‐based oligonucleotide binder) protein. HOB was genetically fused with the enzyme cytochrome P450 BM3, as well as with BMR, the separated reductase domain of BM3. The resulting HOB‐fusion proteins revealed significantly improved rates in ligation with CH‐modified oligonucleotides and DNA origami nanostructures. These results suggest that the efficient self‐assembly of protein‐decorated DNA structures can be greatly improved by fine‐tuning of the electrostatic interactions between proteins and the negatively charged nucleic acid nanostructures.