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从使用度独立 (COIN) 原生质谱学使用定量枪支糖组的绝对亲和关系
Duong T Bui1, James Favell1, Elena N Kitova1
1Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada.
ACS central science
|July 31, 2023
概括
我们开发了度独立 (COIN) -nMS,以从自然库中量化糖结合蛋白 (GBP) 亲和力. 这种方法可以精确测量连接体亲属性,揭示新的特异性和SARS-CoV-2相互作用.
科学领域:
- 分析化学 分析化学
- 生物化学 生化学
- 格莱科米克斯 (Glycomics) 是一个有趣的药物.
背景情况:
- 原生质谱法 (nMS) 对于选糖甘库对糖甘结合蛋白 (GBPs) 有价值.
- 由于自然图书馆中的未知糖甘度,直接亲和度测量具有挑战性.
- 准确的亲和度确定对于理解分子相互作用和连接体发现至关重要.
研究的目的:
- 引入度独立 (COIN) -nMS用于对天然甘氨酸库进行定量选.
- 为了能够直接测量GBP-糖甘相互作用的结合亲和度 (Kd).
- 为了证明COIN-nMS对各种莱克丁和甘氨酸结构的多功能性,包括病毒标.
主要方法:
- 开发了COIN-nMS,利用纳米流电喷气电离辐射器中的受控溶液混合.
- 通过通过nMS分析GBP-甘氨酸复合物的时间依赖的相对丰度变化,量化亲和力 (Kd).
- 验证了COIN-nMS使用纯化甘氨酸和已知的GBP,并将其与对糖化GBP的捕获释放 (CaR) -nMS实现.
主要成果:
- COIN-nMS可靠地确定了已知的GBP-糖甘相互作用的亲和关系.
- COIN-CaR-nMS成功地对复杂的天然甘氨酸库进行了多种类型的 (植物,真菌,病毒,人类) 讲解蛋白的选.
- 通过SARS-CoV-2受体结合域识别了单化杂交和双年性N-glycans的特定识别.
结论:
- COIN-nMS提供了一种可靠的方法,用于从天然甘氨酸库中进行定量联体发现和亲和度测量.
- 该试验揭示了各种单蛋白的细致特异性,并澄清了SARS-CoV-2的结合性质.
- 主体N-甘氨酸的药理性耗尽减少了病毒的进入,表明它们作为附着因子的作用.
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