多重点击选择性tRNA合成酶扩展哺乳动物细胞特异性蛋白质组
Andrew C Yang, Haley du Bois, Niclas Olsson
1Center for Tissue Regeneration, Repair and Restoration , V.A. Palo Alto Healthcare System , Palo Alto , California 94304 , United States.
Journal of the American Chemical Society
|May 19, 2018
概括
研究人员开发了新的生物对角工具,设计了tRNA合成酶的tyrosyl和phenylalanyl,用于精确的哺乳动物细胞标记. 这些工具使得先进的细胞类型特异性蛋白质组学能够更深入地理解生物.
科学领域:
- 生物化学
- 分子生物学
- 蛋白质组学
背景情况:
- 细胞类型特定的蛋白质组对于理解多细胞生物体中的复杂生物过程至关重要.
- 现有的生物直角工具在哺乳动物系统中存在局限性.
研究的目的:
- 开发用于哺乳动物生物直角标记的新型氨基酸-tRNA合成酶.
- 在活哺乳动物中实现细胞类型特定的蛋白质组.
主要方法:
- 设计了一个tyrosyl (ScTyrY43G) 和一个phenylalanyl (MmPheT413G) tRNA合成酶.
- 在新生蛋白质中加入含有亚的非正规氨基酸.
- 用光体或亲属性树脂进行化学选择性标记.
主要成果:
- ScTyrY43G和MmPheT413G都成功标记了人类,仓鼠和老鼠细胞系中的蛋白质.
- 与正规氨基酸相比,突变合成酶可以选择性激活含氨基酸的10倍以上.
- 在小鼠中实现了瘤蛋白质和血分泌物的体内标记.
结论:
- 工程合成酶为哺乳动物残留物特定的生物对角化学提供了新的工具.
- 这些工具在活哺乳动物中促进了更强大和更全面的细胞类型特异性蛋白质组.
- 能够进行高级成像和蛋白质组的质谱特征.
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