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在FLS2-flg22结合接口附近调节酵母糖化,可在诱导ER压力下进行功能性酵母表面显示

Benedikt Dolgikh, Samantha Schulte, Daniel R Woldring

bioRxiv : the preprint server for biology

|December 15, 2025

概括

研究人员为功能植物免疫受体FLS2 (Flagellin Sensing 2) 设计了酵母表面显示. 这种方法优化了FLS2变种的查,以改善植物中的细菌植物病原体检测.

科学领域:

植物免疫力植物免疫力
分子生物学分子生物学
生物技术是生物技术.

背景情况:

像FLAGELLIN SENSING 2 (FLS2) 这样的模式识别受体对于植物对细菌病原体的免疫力至关重要.
在植物中评估FLS2变体是低吞吐量;异质系统需要优化.

研究的目的:

建立一个功能性的酵母表面显示系统,用于Arabidopsis thaliana FLS2 ectodomain.
为了优化FLS2显示器,用于高通量选变种.

主要方法:

在Saccharomyces cerevisiae表面表达的阿拉比多普西斯FLS2ectodomain.
使用突尼卡米辛和热应力进行操纵的N-糖化.
使用流量细胞计和磁珠丰富用于flg22结合确认.
采用结构建模来指导糖甘位点突变发生.

主要成果:

在酵母表面实现了功能FLS2表达和flg22结合.
确定FLS2贩运需要特定的N-糖化.
通过调节糖化和ER应激,证明了flg22结合的改善.
发现的N388变种在压力下增强了功能表现.

结论:

酵母糖化和ER应激反应影响FLS2的功能显示.
开发了一个用于工程酵母表面显示FLS2和相关受体的框架.
这个平台可以实现高通量查,以改善植物病原体检测.

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