在化合物混合物中,一种一般技术来排列蛋白质 - 配体结合的亲缘关系,并确定异质与直接结合位点的竞争
D Allen Annis1, Naim Nazef, Cheng-Chi Chuang
1NeoGenesis Pharmaceuticals Inc., 840 Memorial Drive, Cambridge, MA 02139, USA. aannis@neogenesis.com
Journal of the American Chemical Society
|November 26, 2004
概括
我们开发了一种亲和性选择质谱法 (AS-MS) 方法来排列蛋白质-连接体结合亲和性,并根据结合位点对连接体进行分类. 这个工具通过有效地识别和优化用于疾病标的化合物,有助于药物发现.
科学领域:
- 生物化学 生物化学
- 化学生物学 化学生物学
- 药物发现 药物发现 药物发现
背景情况:
- 组合化学为药物发现提供了巨大的潜力,但需要有效的工具来识别和优化化合物.
- 目前用于蛋白质 - 配体相互作用分析的方法可能是资源密集的,缺乏全面的结合位点分类.
- 开发通用,高效的工具对于利用药物开发中的以多样性为导向的合成至关重要.
研究的目的:
- 引入和验证一种新的亲和力选择-质谱法 (AS-MS) 方法,用于蛋白质-配体亲和力排名和结合位点分类.
- 在药物发现中,为各种蛋白标和配体类提供适用于各种蛋白标的多功能工具.
- 为了能够在不需要复杂的测试的情况下,对结合性亲缘关系和合作性的定量估计.
主要方法:
- AS-MS方法涉及从配体池中选择蛋白质结合化合物的亲和力,使用竞争对手的配体.
- 通过染色学将未结合的连接物从蛋白质-连接物复合物分离出来.
- 通过质谱法 (MS) 分离和识别/定量联体,以确定亲和力和结合部位.
主要成果:
- 证明AS-MS对各种蛋白质类的成功应用,包括激酶 (Akt-1,Zap-70) 和GPCR (M(2) 肌肉性乙胆受体).
- 确定了新的ATP竞争性和全性抑制剂,以及以前未公开的抗剂.
- 验证了该方法在量化估计结合性亲缘关系和结合性合作性方面的能力.
结论:
- AS-MS方法是一种强大的,无标签的技术,用于蛋白质-配体亲和度排名和结合位点分类.
- 它不需要任何化合物或蛋白质修饰,可以在溶液中进行.
- 该方法非常适用于高通量查,优化和药物发现中的结构-活性关系研究.
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