酶生成的硫化的化学发光检测:基质结对H2S对生物醇的选择性产生影响
Journal of the American Chemical Society
|October 8, 2013
概括
新的化学发光硫化物传感器 (CLSS) 能够灵敏地检测硫化 (H2S),这是一个关键的生物调节器. CLSS-2显示出对H2S的高选择性,促进其在生物系统中的测量,并指导未来的传感器设计.
科学领域:
- 生物化学和分子生物学
- 化学生物学 化学生物学
- 分析化学 分析化学
背景情况:
- 硫化 (H2S) 是一种新兴的信号分子,在生理过程和疾病中起着关键作用.
- 由于现有的方法有限,检测内源H2S是具有挑战性的,这阻碍了对其生物功能的理解.
- 现有的检测方法往往缺乏对生物H2S水平所需的灵敏度或选择性.
研究的目的:
- 开发基于反应的新型化学发光硫化物传感器 (CLSS),用于敏感和选择性检测内源H2S.
- 调查CLSS-1和CLSS-2对H2S对其他反应性硫,和氧物种 (RSONS) 的选择性.
- 验证开发的传感器用于测量细胞系统中生物产生的H2S的实用性.
主要方法:
- 合成和描述两个化学发光硫化物传感器,CLSS-1和CLSS-2.
- 对H2S的CLSS-1和CLSS-2的发光响应和检测极限的评估.
- 使用光谱方法对各种RSONS,包括醇 (GSH,Cys,Hcy) 的选择性进行评估.
- 计算DFT研究和NMR实验以阐明差异选择性的机制.
- 应用CLSS-2用于检测由孤立的cystathionine γ-lyase (CSE) 酶和C6细胞产生的H2S.
主要成果:
- 对于H2S,CLSS-1和CLSS-2表现出强烈的发光反应 (128倍和48倍) 和低的检测极限 (0.7 ± 0.3 μM和4.6 ± 2.0 μM).
- 在一系列RSONS中,CLSS-2对H2S具有很高的选择性,包括生物相关的醇.
- CLSS-1和CLSS-2之间的选择性差异归因于基质探针相互作用,影响了阿齐德的可访问性.
- CLSS-2成功地检测到CSE在分离的酶和C6细胞中酶生成的H2S,并区分了活性和抑制状态.
结论:
- CLSS-1和CLSS-2为化学发光H2S的检测和量化提供了敏感和选择性的工具.
- CLSS-2是一种有价值的试剂,用于测量复杂的生物样本中的生物相关H2S水平.
- 对选择性的机械洞察力为设计改进的H2S检测支架提供了基础.
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