N-methylcysteine-mediated total chemical synthesis of ubiquitin thioester.
N-methylcysteine-mediated total chemical synthesis of ubiquitin thioester.
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DOI：
10.1039/c000332h
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发表时间：
2010-05-21
影响因子：
3.2
通讯作者：
Brik A
中科院分区：
文献类型：
--
作者：
Erlich LA;Kumar KS;Haj-Yahya M;Dawson PE;Brik A
泛素硫酯是蛋白质泛素化的关键中间体，通过E1酶以ATP依赖的方式激活泛素的α-COOH而形成。目前用于制备泛素硫酯的方法要么依赖于酶机制，要么依赖于表达蛋白连接技术。本文报道了一种新的化学策略，结合天然化学连接和含n -甲基半胱氨酸肽，首次化学制备泛素硫酯。N-甲基半胱氨酸被用作N→S酰基转移装置，并在其保护形式下作为潜在的硫酯功能。这使我们能够在泛素多肽组装之后通过天然化学连接触发泛素硫酯的形成。合成的泛素硫酯在肽泛素化方面表现出与通过表达获得的泛素硫酯相似的行为。这种方法应该允许在各种泛素化肽和蛋白质的化学操作中具有更高的灵活性，用于结构和生化分析以及泛素链的合成。
Ubiquitin thioester is a key intermediate in the ubiquitylation of proteins and is formed enzymatically through the activation of α-COOH of ubiquitin in an ATP dependent manner using the E1 enzyme. The current methods used for the preparation of ubiquitin thioester rely on either the enzymatic machinery or on expressed protein ligation technology. In this article, we report a new chemical strategy, combining native chemical ligation and N-methylcysteine containing peptides, to chemically prepare ubiquitin thioester for the first time. The N-methylcysteine is utilized as an N→S acyl transfer device, and in its protected form serves as a latent thioester functionality. This enabled us to trigger the formation of ubiquitin thioester subsequent to the assembly of the ubiquitin polypeptide via native chemical ligation. The synthetic ubiquitin thioester showed a similar behavior in peptide ubiquitylation to the one obtained via expression. This approach should allow for higher flexibility in the chemical manipulation of ubiquitin thioester in a wide variety of ubiquitylated peptides and proteins for structural and biochemical analysis and for the synthesis of ubiquitin chains.
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影响因子：
15
作者：
Yan, LZ;Dawson, PE
通讯作者：
Dawson, PE
影响因子：
56.9
作者：
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通讯作者：
KENT, SBH
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作者：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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