Universal guidelines for the conversion of proteins and dyes into functional nanothermometers
Universal guidelines for the conversion of proteins and dyes into functional nanothermometers
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DOI：
10.1002/jbio.201900044
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发表时间：
2019-09-01
影响因子：
2.8
通讯作者：
Thompson, Sebastian A.
中科院分区：
文献类型：
--
作者：
Spicer, Graham;Efeyan, Alejo;Thompson, Sebastian A.
在过去的十年中，化学和光子学的技术进步使得纳米级温度的实时测量成为可能。纳米温度计是专门为传递这些纳米级温度变化而设计的探头，可提供高度的温度、时间和空间分辨率和精度。已经提出了几种不同的方法，包括微型热电偶、基于发光和荧光偏振各向异性的纳米温度计。基于各向异性的纳米温度计在生物相容性方面表现出色，因为它们可以由与染料缀合的内源蛋白构建，从而最大限度地减少任何系统扰动。此外，所得荧光蛋白在进行温度测量时可以保留其天然结构和活性，从而可以从天然环境或任何给定实验系统中的酶促反应期间进行精确的温度记录。为了促进这些纳米温度计未来在研究中的使用，我们在这里提出了一个理论模型，该模型根据蛋白质大小和染料荧光寿命，从任何蛋白质或染料开始预测基于各向异性的温度计的最佳灵敏度。使用该模型，大多数蛋白质和染料都可以转换为纳米温度计。科学界各学科对这些纳米温度计的利用将带来目前技术尚无法获得的新知识和理解。
In the last decade, technological advances in chemistry and photonics have enabled real-time measurement of temperature at the nanoscale. Nanothermometers, probes specifically designed to relay these nanoscale temperature changes, provide a high degree of temperature, temporal, and spatial resolution and precision. Several different approaches have been proposed, including microthermocouples, luminescence and fluorescence polarization anisotropy-based nanothermometers. Anisotropy-based nanothermometers excel in terms of biocompatibility because they can be built from endogenous proteins conjugated to dyes, minimizing any system perturbation. Moreover, the resulting fluorescent proteins can retain their native structure and activity while performing the temperature measurement, allowing precise temperature recordings from the native environment or during an enzymatic reaction in any given experimental system. To facilitate the future use of these nanothermometers in research, here we present a theoretical model that predicts the optimal sensitivity for anisotropy-based thermometers starting with any protein or dye, based on protein size and dye fluorescence lifetime. Using this model, most proteins and dyes can be converted to nanothermometers. The utilization of these nanothermometers by a broad spectrum of disciplines within the scientific community will bring new knowledge and understanding that today remains unavailable with current techniques.