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结构和生化研究产品抑制S-腺甲氨酸合成酶从Corynebacterium glutamicum

Seunghwan Lee¹, Seongmin Kim¹, Il-Kwon Kim²

¹School of Life Sciences, BK21 FOUR KNU Creative BioResearch Group, Kyungpook National University, Daegu 41566, Republic of Korea.

Journal of agricultural and food chemistry

|October 17, 2023

概括

来自Corynebacterium glutamicum的S-adenosylmethionine合成酶 (MetK) 进行了结构分析. 蛋白质工程创造了一个变体 (CgMetK^E68A),可以克服SAM抑制,增强酶活性和稳定性.

科学领域:

生物化学生物化学
结构生物学结构生物学
酶学是一种酶学.

背景情况:

在生物系统中,S-Adenosylmethionine (SAM) 是一个重要的甲基供体.
S-adenosylmethionine合成酶 (MetK) 从甲和ATP合成SAM,使其对细胞功能至关重要.
了解MetK的结构和监管是控制SAM水平的关键.

研究的目的:

为了确定Corynebacterium glutamicumMetK (CgMetK) 在它的apo和结合体结合形式中的晶体结构.
通过其产品SAM.来研究CgMetK的全抑制机制.
设计一种具有减少SAM抑制和改善酶特性的CgMetK变体.

主要方法:

使用X射线结晶学来确定CgMetK的结构.
结构指导蛋白质工程被用来创造突变变体.
在野生类型和人工酶上进行了酶活性测定和热稳定性测试.

主要成果:

晶体结构揭示了CgMetK的活性部位附近的一个全性SAM结合部位.
发现MetK通过竞争性和非竞争性机制被SAM抑制.
该CgMetKE68A变种显示SAM抑制显著降低,酶活性增强.
对CgMetKE68A的结构分析表明,一种新的键破坏了全性SAM结合点,并稳定了酶的四重体形式.

结论:

这项研究阐明了在CgMetK中SAM全抑制的结构基础.
蛋白质工程可以有效地克服产品抑制,从而提高酶功能.
该CgMetKE68A变种代表了一个潜在的有价值的工具,用于生物技术应用需要高效的SAM合成.

关键词:

在Corynebacterium glutamicum的研究中,我们发现这是一种S-adenosylmethionine合成酶.全osteric 抑制的抑制.蛋白质工程工程蛋白质工程

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