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

• Published in: Nature chemistry Vol. 6; no. 9; pp. 804 - 809
• Main Authors: Rosen, Christian B., Kodal, Anne L. B., Nielsen, Jesper S., Schaffert, David H., Scavenius, Carsten, Okholm, Anders H., Voigt, Niels V., Enghild, Jan J., Kjems, Jørgen, Tørring, Thomas, Gothelf, Kurt V.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.09.2014; Nature Publishing Group
• Subjects: 140/58; 639/925/926/1050; Amino Acid Sequence; Analytical Chemistry; Animals; Antibodies; Antibodies - chemistry; Biochemistry; Carrier Proteins - chemistry; Chemistry; Chemistry/Food Science; Deoxyribonucleic acid; DNA; DNA - chemistry; Genetic engineering; Green Fluorescent Proteins; Histidine - chemistry; Humans; Hybridization; Inorganic Chemistry; Labeling; Metals; Metals - metabolism; Molecular biology; Molecular Sequence Data; Organic Chemistry; Physical Chemistry; Proteins; Transferrin - chemistry; Denmark; Aarhus Denmark
• ISSN: 1755-4330; 1755-4349
• DOI: 10.1038/nchem.2003

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 His 6 -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. Conjugation of DNA to proteins often involves a choice between either expressing recombinant proteins with a specific handle, or labelling wild-type proteins with low site-selectivity. Now preorganization of a DNA–ligand complex to a metal-binding site enables site-selective conjugation of a DNA strand to lysine residues of wild-type proteins and antibodies.