可切割的生物素探针用于通过亚酸-基因循环添加来标记生物分子
Janek Szychowski1, Alborz Mahdavi, Jennifer J L Hodas
1Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 East California Boulevard, Pasadena, California 91125, USA.
Journal of the American Chemical Society
|December 15, 2010
概括
新的可切割生物素探针使生物分子的生物对角标记成为可能. 一种新的dialkoxydiphenylsilane (DADPS) 链接探针为蛋白质组学研究提供了高效的裂变和最小质量标记.
科学领域:
- 生物化学 生物化学
- 化学生物学 化学生物学
- 蛋白质组学是指蛋白质组学.
背景情况:
- 亚酸-酸环添加是标记生物分子的关键生物对等反应.
- 可切割探头在蛋白质组学研究中对释放标记分子非常有价值.
- 现有的探头在切割效率或标签大小方面可能存在局限性.
研究的目的:
- 开发和评估用于生物直角标签的新型可切割生物素探针.
- 评估不同连接器设计的裂变效率和特性.
- 确定最佳探针用于蛋白质组学和生物分子研究中的应用.
主要方法:
- 合成了五个可切割的生物素探针,具有不同的链接器化学成分.
- 蛋白质和其他生物分子的生物直角标记使用亚酸-酸环添加.
- 使用不同的化学条件 (减光剂,酸,紫外线照射) 切割标记的生物分子.
- 用绿色光蛋白模型评估一个dialkoxydiphenylsilane (DADPS) 连接探头.
主要成果:
- 五个可切割的生物素探针成功合成和测试.
- 通过各种方法实现了裂变,包括Na2S2O4,2-乙醇,酸,三酸和紫外线.
- DADPS链接探头在0.5小时内以10%的酸有效地进行了切割.
- DADPS探测器表现出选择性结合,并在分裂后留下了一个小的143 Da质量标签.
结论:
- 新型可切割的生物素探针促进了生物对角标记和随后的生物分子释放.
- DADPS链接探头具有高效和选择性,非常适合蛋白质组应用.
- 在基于质谱的研究中,DADPS探头的最小质量标签是有利的.
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