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C-终端域控制烟草细胞中的蛋白质质量和蜘蛛丝的分泌
Chonprakun Thagun1,2, Tomohiro Suzuki2, Yutaka Kodama2,3
1Department of Material Chemistry, Graduate School of Engineering, Kyoto University, Kyoto-Daigaku-Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan.
Advanced biology
|July 6, 2023
概括
研究人员对植物细胞进行了改造,以产生蜘蛛丝蛋白 (MaSp2). 切断特定的蛋白质域显著增加了分泌,为高效的生物聚合物生产铺平了道路.
科学领域:
- 生物技术是生物技术.
- 材料科学 材料科学 材料科学
- 分子生物学分子生物学
背景情况:
- 蜘蛛拖线丝的强度来自主要的囊丝蛋白 (MaSp).
- 完整的MaSp分子是天然纤维线所需的,但生产具有挑战性.
- 之前的努力集中在不同系统的碎片化 MaSp 生产上.
研究的目的:
- 开发一种基于植物细胞的系统,用于整个MaSp2蛋白的细胞外生产.
- 研究影响MaSp2分泌和自我组装成蜘蛛丝纳米纤维的因素.
- 了解 MaSp2 C端域在蛋白质分泌和质量的作用.
主要方法:
- 工程转基因明亮黄色2 (BY-2) 植物细胞系用于分泌的MaSp2表达.
- 将全长的MaSp2与细胞体表达的细胞外产量进行比较.
- 研究了C端域切断对MaSp2分泌水平的影响.
- 评估了产生的MaSp2蛋白质的自我组装特性.
主要成果:
- 转基因BY-2细胞在0.6-1.3微克L-1.1时分泌出全长的MaSp2.
- 细胞外MaSp2产量是细胞内表达的四倍.
- 只有10-15%的分泌的MaSp2被释放到培养介质中.
- 切断C端域增加了分泌量,从0.9到2.8毫克L-1每天.
- 生产的 MaSp2 蛋白质表现出自我组装成蜘蛛丝纳米纤维.
结论:
- 基于植物细胞的系统可以被优化为细胞外生产复合生物聚合物,如蜘蛛丝.
- 马斯普2的C端域在调节蛋白质分泌和质量方面发挥着至关重要的作用.
- 域切断提供了一个有前途的策略,以提高植物表达平台的重组蛋白质产量.
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