Chemical Synthesis of Microtubule-Associated Protein Tau.
Chemical Synthesis of Microtubule-Associated Protein Tau.
复制标题
DOI：
10.1021/jacs.3c07338
复制
发表时间：
2023-10-04
影响因子：
15
通讯作者：
Walczak, Maciej A.
中科院分区：
文献类型：
--
作者：
Powell, Wyatt C.;Jing, Ruiheng;Walczak, Maciej A.
微管相关蛋白Tau（MAPT）的沉积物是称为Tau蛋白病的神经退行性疾病的标志。许多研究已经证明，在诸如阿尔茨海默病（AD）的疾病中，Tau经历广泛的重塑。分布在整个蛋白质序列中的翻译后修饰的附着与临床表现相关。对这些蛋白质改变的系统检查可以揭示它们在健康和疾病状态中的作用。然而，在整个蛋白质链中获得这些修饰的能力是有限的，因为Tau只能重组或通过半合成产生。在这篇文章中，我们描述了Tau的最长2N 4 R同种型的第一个化学合成，由441个氨基酸组成。将2N 4 R Tau分为3个主要片段，总共11个片段，所有片段均通过固相肽合成制备。成功的化学策略依赖于两个半胱氨酸位点（C291和C322）用于天然化学连接（NCL）的策略性使用。这与现代制备蛋白质化学相结合，例如巯基苏氨酸连接（T205）、二硒醚-硒酯连接（D358）和通过均相自由基脱硫将巯基氨基酸突变为天然残基（A40、A77、A119、A157、A246和A390）。合成的成功完成建立了一个强大的和可扩展的路线，以天然蛋白质在数毫克的数量和高纯度。在更广泛的意义上，所提出的策略可以应用于制备其他较短的Tau同种型以及引入所有的翻译后修饰，这些修饰是Tau蛋白病如AD的特征。
Deposits of the microtubule-associated protein Tau (MAPT) serve as a hallmark of neurodegenerative diseases known as tauopathies. Numerous studies have demonstrated that in diseases such as Alzheimer’s disease (AD), Tau undergoes extensive remodeling. The attachment of post-translational modifications distributed throughout the entire sequence of the protein correlates with clinical presentation. A systematic examination of these protein alterations can shed light on their roles in both healthy and diseased states. However, the ability to access these modifications in the entire protein chain is limited as Tau can only be produced recombinantly or through semisynthesis. In this article, we describe the first chemical synthesis of the longest 2N4R isoform of Tau, consisting of 441 amino acids. The 2N4R Tau was divided into 3 major segments and a total of 11 fragments, all of which were prepared via solid-phase peptide synthesis. The successful chemical strategy has relied on the strategic use of two cysteine sites (C291 and C322) for the native chemical ligations (NCLs). This was combined with modern preparative protein chemistries, such as mercaptothreonine ligation (T205), diselenide–selenoester ligation (D358), and mutations of mercaptoamino acids into native residues via homogeneous radical desulfurization (A40, A77, A119, A157, A246, and A390). The successful completion of the synthesis has established a robust and scalable route to the native protein in multimilligram quantities and high purity. In broader terms, the presented strategy can be applied to the preparation of other shorter isoforms of Tau as well as to introduce all post-translational modifications that are characteristic of tauopathies such as AD.
登录
查看更多内容
影响因子：
56.9
作者：
DAWSON, PE;MUIR, TW;KENT, SBH
通讯作者：
KENT, SBH
影响因子：
16.6
作者：
Ansaloni, Annalisa;Wang, Zhe-Ming;Lashuel, Hilal A.
通讯作者：
Lashuel, Hilal A.
影响因子：
4.8
作者：
Boyko, Solomiia;Surewicz, Witold K
通讯作者：
Surewicz, Witold K
DOI：
10.3390/molecules25245789
发表时间：
2020-12-08
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Breijyeh Z;Karaman R
通讯作者：
Karaman R
影响因子：
5.5
作者：
Landrieu, Isabelle;Dupre, Elian;Sinnaeve, Davy;El Hajjar, Lea;Smet-Nocca, Caroline
通讯作者：
Smet-Nocca, Caroline