具有高度选择性的离线光探测器,用于在活细胞中对硫素减少酶进行成像
Liangwei Zhang1, Dongzhu Duan, Yaping Liu
1State Key Laboratory of Applied Organic Chemistry and College of Chemistry and Chemical Engineering, Lanzhou University , Lanzhou, Gansu 730000, China.
Journal of the American Chemical Society
|December 20, 2013
概括
研究人员开发了TRFS-green,这是第一个用于哺乳动物硫素减少酶 (TrxR) 的光探针. 这种探测器可以选择性地检测和成像活细胞中的TrxR活性,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
- 分子成像学分子成像学
背景情况:
- 铁素还原酶 (TrxR) 是细胞氧化还原恒温的关键酶.
- TrxR的失调与包括癌症在内的各种疾病有关.
- 需要有选择性和敏感的探针来研究生物系统中的TrxR活性.
研究的目的:
- 设计,合成和评估一种新型的光探测器,用于发现哺乳动物的硫素减少酶 (TrxR).
- 为了证明探头的选择性,灵敏性和在成像中有用的TrxR活动在体外和活细胞中.
- 探索探针在开发TrxR抑制剂的基于细胞的查试验方面的潜力.
主要方法:
- 设计和合成TRFS-green,一个光探针与1,2-dithiolane支架和灭的纳夫他林胺.
- 在体外评估探针对相关酶和硫醇的选择性.
- 高性能液态色谱 (HPLC) 分析以确认TrxR介导的转化.
- 细胞测试以将光信号与内源的TrxR活性相关联.
- 活细胞成像实验可视化TrxR活动并评估抑制剂效应.
主要成果:
- 在TrxR介导的切割过程中,TRFS绿色表现出一个关闭的光反应.
- 探针显示TrxR对其他还原酶和醇具有很高的选择性.
- 细胞提取物中的光信号与TrxR活性直接相关.
- 在活细胞中成功对TrxR活性进行成像.
- 观察到TrxR抑制剂对光信号的剂量依赖抑制.
结论:
- TRFS-green是第一个针对哺乳动物TrxR.R的选择性光探针.
- 该探测器能够实时成像活细胞中的TrxR活动.
- TRFS-green促进了对TrxR抑制剂的选试验的开发,这可能导致新的治疗药物.
- 这种探测器将有助于进一步了解TrxR的生理和病理生理作用.
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