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在一个活细胞中基于电化学分子陷的低丰度酶的检测

Rongrong Pan¹, Dengchao Wang², Kang Liu¹

¹The State Key Lab of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, Jiangsu, P. R. China.

Journal of the American Chemical Society

|September 16, 2022

概括

研究人员开发了一种新的电化学分子陷,用于测量单个活细胞内的低丰度酶活性. 这一突破使得我们能够精确地检测诸如菌酶之类的酶,

科学领域:

生物化学
分析化学
细胞生物学

背景情况:

在单个细胞中测量低丰度酶活性对于理解生物功能至关重要.
现有的方法难以检测活细胞中的微量酶.

研究的目的:

开发一种超敏感的电化学策略,以检测和量化活细胞中单个分子水平的低丰度酶活性.
克服当前技术在本地细胞环境中测量酶活性的局限性.

主要方法:

在纳米管尖上设计了一种电化学分子陷,利用电流 (EOF) 来限制酶.
酶被捕获以持续催化基质转化,产生可检测的过氧化副产品.
使用电阻脉冲传感来验证检测灵敏度,低至15个酶分子.

主要成果:

这项研究成功地测量了约60个无刺激的J774细胞中的基酶分子的活性.
这种检测水平以前是现有的方法无法实现的.
该技术表现出高灵敏度,有效检测脂质体中的15个分子.

结论:

开发的电化学分子陷可以在活细胞中检测单个分子的电化学反应.
这种方法解决了在生理条件下研究低丰度蛋白质活性的挑战.
这些发现显著提升了对细胞行为中的蛋白质作用的理解.

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