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从in-silico查到in-vitro评估:通过工程PP1突变变种增强微囊的检测能力

Ezequiel J Alba-Posse¹, Carlos David Bruque², Yamila Gándola³

¹Facultad de Ciencias Exactas y Naturales, Departamento de Fisiología y Biología Molecular y Celular, Instituto de Biociencias, Biotecnología y Biología Traslacional (iB3), Universidad de Buenos Aires, Intendente Güiraldes 2160, C1428EGA Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQB Buenos Aires, Argentina.

Journal of structural biology

|November 7, 2023

概括

研究人员增强了一种蛋白质酸酶1 (PP1) 变体,p.D197F,以改善微囊素 (MC) 检测. 这种突变显示了MC的抑制增加,为有害藻类繁殖提供了更敏感的生物传感器.

科学领域:

生物技术是生物技术.
生物化学生物化学
环境科学环境科学

背景情况:

有害的藻类繁殖 (HABs) 产生毒素,造成全球健康风险.
微囊 (MCs) 是一种具有致癌性和肝毒性作用的普遍的毒素.
蛋白质酸酶抑制试验 (PPIAs) 对MC检测具有成本效益.

研究的目的:

设计和评估蛋白质酸酶1 (PP1) 的突变变体.
增强PP1作为微囊素 (MC) 生物传感器的能力.
为了提高MC检测的灵敏度和可靠性.

主要方法:

在PP1.1中对活性位点和突变发生的内选.
稳定性和对接亲和度计算使用MC-LR 毒素.
选择PP1突变变种的体外表达和评估.

主要成果:

确定了19个有前途的PP1突变,其亲和力和稳定性得到改善.
四种突变 (p.D197F,p.Q249Y,p.S129W,p.D220Q) 被选择用于体外测试.
突变p.D197F与野生类型PP1.1相比,显示了MC-LR显著增加的抑制.

结论:

这种p.D197F PP1变种表现出微囊素增强的抑制作用.
这种工程变体具有实际MC检测应用的潜力.
改进的PP1变种可以导致更有效的毒素监测工具.

关键词:

生物传感器是一种生物传感器.蓝色毒素 (Cyanotoxins) 是一种有毒物质.停靠对接停靠对接蛋白质工程是一种蛋白质工程.谢林/三氨酸酸酶的作用.

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