期刊
CHEMBIOCHEM
卷 17, 期 12, 页码 1102-1106出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201600039
关键词
bioconjugates; DNA nanostructures; enzymes; self-assembly
资金
- Deutsche Forschungsgemeinschaft [Ni399/10]
- Helmholtz programme BioInterfaces in Technology and Medicine
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 P450BM3, 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.
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