增强赖氨酸衍生物与非天然氨基酸甲酯形式的蛋白质结合。
Enhancing the incorporation of lysine derivatives into proteins with methylester forms of unnatural amino acids.
机构信息
Department of Chemistry, Center for Diagnostics and Therapeutics, Georgia State University, Atlanta, GA 30302, United States.
Department of Chemistry, Virginia Commonwealth University, Richmond, VA 23284, United States.
出版信息
Bioorg Med Chem Lett. 2020 Jan 15;30(2):126876. doi: 10.1016/j.bmcl.2019.126876. Epub 2019 Dec 4.
Abstract
We have improved the incorporation of l- and d-forms of unnatural amino acid (UAA) N-thiaprolyl-l-lysine (ThzK) into ubiquitin (UB) and green fluorescent protein (GFP) by 2-6 folds with the use of the methylester forms of the UAAs in E coli cell culture. We also improved the yields of UAA-incorporated UB and GFP with the methylester forms of N-Boc-l-Lysine (BocK) and N-propargyl-l-Lysine (PrK) by 2-5 folds compared to their free acid forms. Our work demonstrated that using methylester-capped UAAs for protein expression is a useful strategy to enhance the yields of UAA-incorporated proteins.
摘要
我们通过在大肠杆菌细胞培养中使用非天然氨基酸（UAA）N-硫代脯氨酰-L-赖氨酸（ThzK）甲酯形式，将 L-和 D-型非天然氨基酸（UAA）的掺入提高了 2-6 倍，从而提高了泛素（UB）和绿色荧光蛋白（GFP）的掺入效率。与游离酸形式相比，我们还使用 N-Boc-L-赖氨酸（BocK）和 N-炔丙基-L-赖氨酸（PrK）的甲酯形式，将 BocK 和 PrK 掺入 UB 和 GFP 的产率提高了 2-5 倍。我们的工作表明，使用甲酯封端的 UAA 进行蛋白质表达是提高 UAA 掺入蛋白产量的有效策略。
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