苯丙氨酸
叠氮化物
遗传密码
化学
氨基酸
蛋白质生物合成
融合蛋白
产量(工程)
生物化学
基因
重组DNA
有机化学
材料科学
冶金
作者
Xiaofeng Ma,Bing Wei,Enlin Wang
标识
DOI:10.1016/j.pep.2022.106158
摘要
Azide, the most used photo-crosslinking group, facilitates the analysis of protein structure and function. This group is particularly useful when photochemically label antibodies and examine protein-protein interactions. The use of the expanded genetic code technique allows the special labeling of the functional azide group in proteins by adding the unnatural amino acid (UAA), p-azido-l-phenylalanine (AzF), in response to the amber codon during translation. However, a low UAA uptake rate due to mass transfer resistance in the cell membrane may lead to the early termination of the full-length protein. This study reports a general method for the efficient in vivo incorporation of AzF into the target protein by improving cell permeability using organic solvents. As expected, the yield of the full-length protein was significantly increased, which indicated that the AzF uptake was greatly improved due to the addition of organic solvents. Our method can serve as a good reference for improving the genetic incorporation of other kinds of UAAs into proteins.
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