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Nat Chem. 2014 Sep;6(9):804-9. doi: 10.1038/nchem.2003. Epub 2014 Jul 20.

Template-directed covalent conjugation of DNA to native antibodies, transferrin and other metal-binding proteins.

Author information

1
1] Center for DNA Nanotechnology and Interdisciplinary Nanoscience Center, Gustav Wieds Vej 14, 8000 Aarhus C, Denmark [2] Department of Chemistry, Langelandsgade 140, 8000 Aarhus C, Denmark [3].
2
1] Center for DNA Nanotechnology and Interdisciplinary Nanoscience Center, Gustav Wieds Vej 14, 8000 Aarhus C, Denmark [2] Department of Molecular Biology and Genetics, Aarhus University, C. F. Møllers Allé 3, 8000 Aarhus C, Denmark.
3
Department of Molecular Biology and Genetics, Science Park, Aarhus University, Gustav Wieds Vej 10C, 8000 Aarhus C, Denmark.
4
1] Center for DNA Nanotechnology and Interdisciplinary Nanoscience Center, Gustav Wieds Vej 14, 8000 Aarhus C, Denmark [2] Department of Chemistry, Langelandsgade 140, 8000 Aarhus C, Denmark.

Abstract

DNA-protein conjugates are important in bioanalytical chemistry, molecular diagnostics and bionanotechnology, as the DNA provides a unique handle to identify, functionalize or otherwise manipulate proteins. To maintain protein activity, conjugation of a single DNA handle to a specific location on the protein is often needed. However, preparing such high-quality site-specific conjugates often requires genetically engineered proteins, which is a laborious and technically challenging approach. Here we demonstrate a simpler method to create site-selective DNA-protein conjugates. Using a guiding DNA strand modified with a metal-binding functionality, we directed a second DNA strand to the vicinity of a metal-binding site of His6-tagged or wild-type metal-binding proteins, such as serotransferrin, where it subsequently reacted with lysine residues at that site. This method, DNA-templated protein conjugation, facilitates the production of site-selective protein conjugates, and also conjugation to IgG1 antibodies via a histidine cluster in the constant domain.

PMID:
25143216
DOI:
10.1038/nchem.2003
[Indexed for MEDLINE]

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