用化学蛋白质学方法分析NAD相互作用体
Justina Šileikytė1, Sunil Sundalam1, Larry L David1
1Department of Chemical Physiology and Biochemistry, Oregon Health and Science University, Portland, Oregon 97239, United States.
Journal of the American Chemical Society
|April 29, 2021
概括
研究人员开发了新的化学探针,以识别与尼古丁胺二核酸 (NAD+) 相互作用的蛋白质. 这些工具有助于发现新的NAD+结合蛋白,扩大我们对细胞功能的理解.
科学领域:
- 生物化学
- 化学生物学
- 蛋白质组学
背景情况:
- 尼古丁胺氨基二核酸 (NAD+) 对于新陈代谢至关重要,并作为聚ADP- 核糖聚合酶 (PARP) 等酶的基质.
- 由于缺乏有效的化学工具,NAD+相互作用的全部范围,称为NAD交互体,可能被低估.
研究的目的:
- 设计,合成和验证可点击的新型光亲和标记 (PAL) 探针,用于描述NAD+结合蛋白.
- 使用这些化学工具识别NAD相互作用的新组件.
主要方法:
- 基于NAD+的可点击PAL探针,特别是2-和6-ad-BAD的开发.
- 在化学蛋白质组学实验中应用探针来识别NAD+结合蛋白.
- 通过2-ad-BAD探针对蛋白质进行紫外线依赖的标记.
主要成果:
- 2-ad-BAD探测器证明了有效的,依赖于紫外线的PARP标签.
- 使用2和6-ad-BAD的蛋白质组学研究确定了以前已知的和新的NAD+/NADH结合蛋白.
- 验证探测器在探测NAD相互作用中的实用性.
结论:
- 开发的可点击的PAL探针,2-和6-ad-BAD,是研究NAD相互作用的有效工具.
- 这些探测器有助于发现新的NAD+结合蛋白,进步对细胞过程中的NAD+作用的理解.
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