DNA-Templated Introduction of an Aldehyde Handle in Proteins
详细信息 查看全文
- 作者：Dr. Anne Louise B. Kodal ; Dr. Christian B. Rosen ; Michael R. Mortensen ; Dr. Thomas Tø ; rring and Prof. Kurt V. Gothelf
- 刊名：ChemBioChem
- 出版年：2016
- 出版时间：July 15, 2016
- 年：2016
- 卷：17
- 期：14
- 页码：1338-1342
- 全文大小：838K
- ISSN：1439-7633
文摘
Many medical and biotechnological applications rely on protein labeling, but a key challenge is the production of homogeneous and site-specific conjugates. This can rarely be achieved by simple residue-specific random labeling, but generally requires genetic engineering. Using site-selective DNA-templated reductive amination, we created DNA–protein conjugates with control over labeling stoichiometry and without genetic engineering. A guiding DNA strand with a metal-binding functionality facilitates site-selectivity by directing the coupling of a second reactive DNA strand in the vicinity of a protein metal-binding site. We demonstrate DNA-templated reductive amination for His6-tagged proteins and metal-binding proteins, including IgG1 antibodies. We also used a cleavable linker between the DNA and the protein to remove the DNA and introduce a single aldehyde on the protein. This functions as a handle for further modifications with desired labels. In addition to directing the aldehyde positioning, the DNA provides a straightforward route for purification between reaction steps.