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在C-C键形成后捕获一个多基酸合成模块,揭示了过渡性酸载体域相互作用

Maria Dell¹, Mai Anh Tran², Michael J Capper³

¹Department of Biomolecular Chemistry, Leibniz Institute for Natural Product Research and Infection Biology (HKI), 07745, Jena, Germany.

Angewandte Chemie (International ed. in English)

|December 22, 2023

概括

研究人员研究了模块化聚基酸合成酶 (PKSs),在C-C键形成后揭示了关键的域间相互作用. 这为未来的工程提供了对这些大型酶复合物的动态的洞察力.

科学领域:

生物化学生物化学
结构生物学结构生物学
分子生物学分子生物学

背景情况:

模块化多基合成酶 (PKSs) 是大型的酶复合体,负责合成各种生物活性化合物.
在PKS中,从乙载体蛋白 (ACP) 到合成酶 (KS) 域的基质传递的精确机制尚未完全理解.
了解这些动态对于设计PKS用于新型化合物生产至关重要.

研究的目的:

阐明根素PKS的链分支模块中的结构动态和域间相互作用.
为了研究基质到合成酶 (KS) 域的输送机制.
为模块化PKS装配线的合理设计提供见解.

主要方法:

基于机制的交叉连接使用合成基质替代品.
电子显微镜 (cryo-EM) 用于确定交联复合体的结构.
AlphaFold2预测用于识别潜在的结合部位.
核磁共振 (NMR) 谱学用于研究溶液状态复合物的形成.
独立领域的光化学交联/质谱学.

主要成果:

使用冷EM确定了C-C键形成后二元蛋白质复合物的不对称状态.
AlphaFold2的预测表明了两个不同的乙载体蛋白 (ACP) 结合点,包括一个潜在的基质加载点.
核磁共振和交叉链接/质谱学证实了溶液中的过渡性,链接器独立的域间相互作用.

结论:

在C-C键形成后捕获的分支PKS模块的结构洞察力增强了对PKS域动态的理解.
这些发现揭示了对基质加工至关重要的关键域间相互作用.
这项工作为PKS系统的未来工程提供了宝贵的信息.

关键词:

乙烯基载体蛋白质乙烯基载体蛋白质生物合成生物合成交叉连接 (crosslinking) 是一种交叉连接.电子显微镜电子显微镜模块化多基化合成酶

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