The Chemical Synthesis of Site-Specifically Modified Proteins Via Diselenide-Selenoester Ligation.

The Chemical Synthesis of Site-Specifically Modified Proteins Via Diselenide-Selenoester Ligation.
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通过二硒化物-硒酯连接化学合成位点特异性修饰的蛋白质。

DOI:
10.1007/978-1-0716-1617-8_18
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发表时间:
2021
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
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通讯作者:
Griffiths RC
Griffiths RC
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作者:
Griffiths RC

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肽连接技术使得能够控制天然和工程化蛋白质的化学合成,包括显示位点特异性翻译后修饰(PTM)和非蛋白质功能性的实例。二硒醚-硒酯连接(DSL)是最近添加到合成方法中的,其提供了优于现有策略的几个优点。标准DSL反应涉及携带N-末端硒代半胱氨酸(Sec)残基的肽与携带C-末端硒代酯的片段的无添加剂连接。这种操作简单的连接在空间位阻连接处快速进行,并且在宽pH范围内有效。将脱硒和氧化脱硒技术并入DSL方案中使得连接位点处的Sec残基能够分别转化为丙氨酸(Ala)和丝氨酸(Ser),从而增强了该方法的范围和通用性。在这一章中,我们描述了DSL的应用一锅化学合成蛋白质通过两个组件和三个组件连接途径。
Peptide ligation techniques enable the controlled chemical synthesis of native and engineered proteins, including examples that display site-specific post-translational modifications (PTMs) and non-proteinogenic functionality. Diselenide-selenoester ligation (DSL) is a recent addition to the synthetic methodology that offers several advantages over existing strategies. The standard DSL reaction involves the additive-free ligation of a peptide carrying an N-terminal selenocysteine (Sec) residue with a fragment bearing a C-terminal selenoester. This operationally simple ligation proceeds rapidly at sterically hindered junctions and is efficient across a broad pH range. The incorporation of deselenization and oxidative deselenization techniques into the DSL protocol enables conversion of the Sec residue at the ligation site to alanine (Ala) and serine (Ser), respectively, thus enhancing the scope and versatility of the method. In this chapter, we describe the application of DSL to the one-pot chemical synthesis of proteins via both two-component and three-component ligation pathways.
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