Kim Sanggil, Ko Wooseok, Sung Bong Hyun, Kim Sun Chang, Lee Hyun Soo
Department of Chemistry, Sogang University, Seoul 121-742, Republic of Korea.
Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea.
Bioorg Med Chem. 2016 Nov 15;24(22):5816-5822. doi: 10.1016/j.bmc.2016.09.035. Epub 2016 Sep 15.
Proteins often function as complex structures in conjunction with other proteins. Because these complex structures are essential for sophisticated functions, developing protein-protein conjugates has gained research interest. In this study, site-specific protein-protein conjugation was performed by genetically incorporating an azide-containing amino acid into one protein and a bicyclononyne (BCN)-containing amino acid into the other. Three to four sites in each of the proteins were tested for conjugation efficiency, and three combinations showed excellent conjugation efficiency. The genetic incorporation of unnatural amino acids (UAAs) is technically simple and produces the mutant protein in high yield. In addition, the conjugation reaction can be conducted by simple mixing, and does not require additional reagents or linker molecules. Therefore, this method may prove very useful for generating protein-protein conjugates and protein complexes of biochemical significance.
蛋白质通常与其他蛋白质结合形成复杂结构发挥功能。由于这些复杂结构对于复杂功能至关重要，开发蛋白质-蛋白质缀合物已引起研究兴趣。在本研究中，通过将含叠氮基的氨基酸基因掺入一种蛋白质，将含双环壬炔（BCN）的氨基酸基因掺入另一种蛋白质，实现了位点特异性蛋白质-蛋白质缀合。对每种蛋白质中的三到四个位点进行缀合效率测试，三种组合显示出优异的缀合效率。非天然氨基酸（UAA）的基因掺入技术简单，且能高产率产生突变蛋白。此外，缀合反应可通过简单混合进行，无需额外试剂或连接分子。因此，该方法对于生成具有生化意义的蛋白质-蛋白质缀合物和蛋白质复合物可能非常有用。
