Protease probes built from DNA: multispectral fluorescent DNA-peptide conjugates as caspase chemosensors.
Protease probes built from DNA: multispectral fluorescent DNA-peptide conjugates as caspase chemosensors.
复制标题
DOI：
10.1002/anie.201007805
复制
发表时间：
2011-05-23
期刊：
影响因子：
--
通讯作者：
Kool ET
中科院分区：
文献类型：
--
作者：
Dai N;Guo J;Teo YN;Kool ET
酶活性的直接监测在许多领域都有广泛的用途，从生物化学到药物化学和生物学一个生物过程通常涉及多种酶，它们可以单独或协同工作来控制特定的生物事件。监测它们的活性可以为这一过程的进展提供标记（如凋亡中发生的caspase活性级联），并提供有关相互作用物种的机制和时间的信息。在单一事件或不同过程中同时跟踪多个目标的能力可以极大地提高我们的基本认识，并促进生物学和临床研究。然而，由于现有荧光团的特性，酶的实时荧光多色传感面临一些技术限制。不同颜色的传统有机荧光团具有不同的吸收波长，需要不同的滤光片来监测每个物种。此外，使用不同的荧光团可能需要开发不同的共轭策略，以及复杂的荧光团-猝灭剂对选择。解决这些问题的一个潜在方法是使用无机量子点（QDs），它允许单波长激发，并且可以产生大小和成分可调的发射然而，在均匀化学修饰和细胞递送方面的困难，以及它们相对较大的尺寸（15-35 nm）和细胞毒性[3]，给它们在传感领域的应用带来了一些限制，特别是在细胞环境中。
The direct monitoring of enzyme activities is broadly useful in many fields, ranging from biochemistry to medicinal chemistry and biology.[1] A biological process often involves multiple enzymes, which can work independently or cooperatively to control a specific biological event. Monitoring their activities can provide markers of the progress of such a process (such as the cascade of caspase activity that occurs in apoptosis), and yield information about the mechanism and timing of the interacting species. The ability to track multiple targets in a single event or different processes simultaneously could greatly improve our basic understanding, and facilitate biological and clinical studies as well.However, the real-time fluorescent multi-color sensing of enzymes faces some technical limitations due to the properties of available fluorophores. Traditional organic fluorophores in distinct colors have disparate absorption wavelengths, requiring different filter sets for monitoring each species. Moreover, the use of different fluorophores can require the development of different conjugation strategies, and complicated fluorophore–quencher pair selection. One potential approach to addressing these problems is the use of inorganic quantum dots (QDs), which allow single-wavelength excitation, and can generate size-and composition-tunable emissions.[2] However, difficulties in uniform chemical modification and cellular delivery, along with their relatively large size (15–35 nm) and cytotoxicity [3] present some limitations of their own for application in sensing, especially in the cellular context.