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来自酵母,线虫和人类的多利霍尔激酶可以相互替代,并交换它们的域,在酵母中产生活性化学酶

Danguole Ziogiene¹, Andrius Burdulis¹, Albertas Timinskas¹

¹Institute of Biotechnology, Life Sciences Center, Vilnius University, Vilnius, Lithuania.

|November 7, 2024

概括

多利霍尔酶 (DK) 有两个域对蛋白质糖化至关重要. C端域对活动至关重要,而N端域可以调节它,使物种之间功能互补成为可能.

科学领域:

生物化学生化学
分子生物学分子生物学
酵母遗传学酵母遗传学

背景情况:

蛋白质糖化对真核细胞功能至关重要.
多利霍尔酶 (DK) 酸化多利霍尔,这是糖化中的关键步骤.
克劳维罗米切斯乳糖DK突变体对多利胆酸盐缺乏症有耐受性.

研究的目的:

研究多利胆酶 (DK) 的结构和功能领域.
确定DK的N端和C端域在蛋白质糖化中的作用.
评估不同物种的DK领域的功能补充.

主要方法:

对DK同类的比较序列分析.
在DK的AlphaFold3结构建模.
在Kluyveromyces lactis和Saccharomyces cerevisiae突变体中的DK域的功能分析.
构建和测试仿真DK酶.

主要成果:

预计DK将有两个结构/功能领域.
C端域对DK活动至关重要,但单独对K. lactis的生存不够.
过度表达N-或C-终端域,或共同表达,部分恢复了糖化.
来自人类和C. elegans的异质N端域补充了酵母DK功能.
由K. lactis和C. elegans的N端域形成的化学酶是活跃的.

结论:

DK的C端域对其酶活性至关重要.
N-终端域起着调节作用,可以在物种之间进行功能替代.
域相互作用对于维持足够的蛋白质糖化水平至关重要.

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