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基于喹啉的光不稳定保护策略促进蛋白质的高效组装
作者:小柯机器人 发布时间:2022/1/21 16:38:47

上海交通大学王平团队报道了基于喹啉的光不稳定保护策略促进蛋白质的高效组装。相关研究成果于2022年1月16日发表在《美国化学会杂志》。

天然化学连接(NCL)为组装具有精确化学特征的蛋白质提供了一个强有力的解决方案,从而能够详细研究蛋白质的结构-功能关系。作为NCL的延伸,脱硫和表达蛋白连接(EPL)技术的发现极大地扩展了通过化学连接获得大型或具有挑战性的蛋白质序列的有效途径。尽管其优越的可靠性,NCL脱硫协议需要正交保护策略,以能够在天然Cys存在的情况下进行选择性脱硫,这对其合成应用至关重要。与传统的硫醇保护基团相比,光不稳定保护基团(PPGs)在辐照后被去除,简化了蛋白质组装,因此对肽支架的扰动最小。然而,目前的PPG策略主要局限于硝基苄基衍生物,其NCL脱硫不兼容。

该文中,研究人员首次提出基于喹啉的半胱氨酸PPG可促进各种连接策略,包括迭代NCL和EPL脱硫方法。蛋白质序列中多个半胱氨酸残基的7-(哌嗪-1-基)-2-(甲基)喹啉基(PPZQ)CAG可通过后期修饰容易引入,而PPZQ在水凝胶缓冲液中通过光解可以高效地无迹去除。此外,PPZQ基团与自由基脱硫相容。通过一锅迭代连接法和EPL脱硫法合成γ-突触核蛋白和磷酸化半胱氨酸蛋白酶抑制剂-S突出了该策略的效率。此外,对表达的白介素-34片段的成功的六倍保护和去保护表明了该策略在蛋白质笼化/脱笼研究中的巨大潜力。

附:英文原文

Title: Quinoline-Based Photolabile Protection Strategy Facilitates Efficient Protein Assembly

Author: Siyao Wang, Qingqing Zhou, Yunxue Li, Bingcheng Wei, Xinliang Liu, Jie Zhao, Farong Ye, Zhongneng Zhou, Bei Ding, Ping Wang

Issue&Volume: January 16, 2022

Abstract: Native chemical ligation (NCL) provides a powerful solution to assemble proteins with precise chemical features, which enables a detailed investigation of the protein structure–function relationship. As an extension to NCL, the discovery of desulfurization and expressed protein ligation (EPL) techniques has greatly expanded the efficient access to large or challenging protein sequences via chemical ligations. Despite its superior reliability, the NCL-desulfurization protocol requires orthogonal protection strategies to allow selective desulfurization in the presence of native Cys, which is crucial to its synthetic application. In contrast to traditional thiol protecting groups, photolabile protecting groups (PPGs), which are removed upon irradiation, simplify protein assembly and therefore provide minimal perturbation to the peptide scaffold. However, current PPG strategies are mainly limited to nitro-benzyl derivatives, which are incompatible with NCL-desulfurization. Herein, we present for the first time that quinoline-based PPG for cysteine can facilitate various ligation strategies, including iterative NCL and EPL-desulfurization methods. 7-(Piperazin-1-yl)-2-(methyl)quinolinyl (PPZQ) caging of multiple cysteine residues within the protein sequence can be readily introduced via late-stage modification, while the traceless removal of PPZQ is highly efficient via photolysis in an aqueous buffer. In addition, the PPZQ group is compatible with radical desulfurization. The efficiency of this strategy has been highlighted by the synthesis of γ-synuclein and phosphorylated cystatin-S via one-pot iterative ligation and EPL-desulfurization methods. Besides, successful sextuple protection and deprotection of the expressed Interleukin-34 fragment demonstrate the great potential of this strategy in protein caging/uncaging investigations.

DOI: 10.1021/jacs.1c10324

Source: https://pubs.acs.org/doi/10.1021/jacs.1c10324

 

期刊信息

JACS:《美国化学会志》,创刊于1879年。隶属于美国化学会,最新IF:14.612
官方网址:https://pubs.acs.org/journal/jacsat
投稿链接:https://acsparagonplus.acs.org/psweb/loginForm?code=1000