协同生物对等标记揭示了内源性转换性O-GlcNAc修饰的新生蛋白
Yanping Zhu1,2, Lianne I Willems1, Daniela Salas1,3
1Department of Chemistry, Simon Fraser University, Burnaby, British Columbia V5A 1S6, Canada.
Journal of the American Chemical Society
|September 2, 2020
概括
这项研究确定了O-GlcNAc在翻译过程中修饰的内源蛋白质,揭示了蛋白质平衡的新机制. 这些发现有助于我们更好地了解O-GlcNAc
科学领域:
- 生物化学
- 分子生物学
- 细胞生物学
背景情况:
- 对于细胞生理和健康,O-GlcNAc对氨酸/氨酸残留物的修饰至关重要.
- O-GlcNAc正在成为细胞蛋白质平衡的关键调节剂.
- 之前的研究表明O-GlcNAc的修饰可以通过转换发生,从而防止新生的多降解.
研究的目的:
- 开发标记和识别内源性O-GlcNAc修饰的新生多链的策略.
- 研究共翻译O-GlcNAc在蛋白质质量控制和蛋白质稳定中的作用.
- 识别经过共翻译的O-GlcNAc修饰的特定内源蛋白.
主要方法:
- 使用O-propargyl-puromycin (OPP) 标记新生链的双重代谢工程.
- 使用四氧化乙烯-2-N-酸乙烯-2-脱氧-d-甲酸 (Ac4GalNAz) 的代谢糖类工程以准O-GlcNAc.
- 用于强大的标签的序列化选择性结合策略.
- 两步丰富的糖基化新生链,然后进行射门蛋白质组.
主要成果:
- 成功标记和识别内源性转换O-GlcNAc修饰的蛋白质
- 对三种已识别的蛋白质进行协译O-GlcNAcylation的验证.
- 从复杂混合物中分离低丰度内源分析物的策略演示.
结论:
- 开发了新的方法来隔离和识别转换O-GlcNAc修饰的蛋白质.
- 提供了O-GlcNAc在细胞质蛋白质质量控制中的作用.
- 开辟了研究协译蛋白修饰在蛋白质稳定中的更广泛作用的途径.
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