应用硒代半胱氨酸介导的表达蛋白连接技术制备膜酶硒蛋白 S。
Applying selenocysteine-mediated expressed protein ligation to prepare the membrane enzyme selenoprotein S.
机构信息
Department of Chemistry and Biochemistry, University of Delaware, Newark, DE, United States.
Department of Chemistry and Biochemistry, University of Delaware, Newark, DE, United States; Department of Pharmaceutical Chemistry, University of California, San Francisco, CA, United States.
出版信息
Methods Enzymol. 2022;662:159-185. doi: 10.1016/bs.mie.2021.10.023. Epub 2021 Dec 24.
The intrinsically disordered membrane-bound selenoprotein s (selenos) takes part in the protein quality control pathway, vesicle trafficking, and NF-kB signaling. The reactive selenocysteine (Sec) at the penultimate position is responsible for its enzymatic activity. We report the preparation of the soluble segment as well as the full-length selenos using expressed protein ligation. This chapter discusses the practical considerations of expressed protein ligation using selenopeptides and describes our optimized procedure for the semi-synthesis of selenos.
固有无序的膜结合硒蛋白 s（selenos）参与蛋白质质量控制途径、囊泡运输和 NF-kB 信号转导。倒数第二位的反应性硒代半胱氨酸 (Sec) 负责其酶活性。我们使用表达蛋白连接法制备了可溶性片段和全长 selenos。本章讨论了使用硒肽进行表达蛋白连接的实际考虑因素，并描述了我们优化的半合成 selenos 的程序。
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