Roeffaers Maarten B J, De Cremer Gert, Uji-i Hiroshi, Muls Benîot, Sels Bert F, Jacobs Pierre A, De Schryver Frans C, De Vos Dirk E, Hofkens Johan
Department of Microbial and Molecular Systems, Centre for Surface Chemistry and Catalysis, Katholieke Universiteit Leuven, Kasteelpark Arenberg 23, B-3001 Leuven, Belgium.
Proc Natl Acad Sci U S A. 2007 Jul 31;104(31):12603-9. doi: 10.1073/pnas.0610755104. Epub 2007 Jul 30.
The ever-improving time and space resolution and molecular detection sensitivity of fluorescence microscopy offer unique opportunities to deepen our insights into the function of chemical and biological catalysts. Because single-molecule microscopy allows for counting the turnover events one by one, one can map the distribution of the catalytic activities of different sites in solid heterogeneous catalysts, or one can study time-dependent activity fluctuations of individual sites in enzymes or chemical catalysts. By experimentally monitoring individuals rather than populations, the origin of complex behavior, e.g., in kinetics or in deactivation processes, can be successfully elucidated. Recent progress of temporal and spatial resolution in single-molecule fluorescence microscopy is discussed in light of its impact on catalytic assays. Key concepts are illustrated regarding the use of fluorescent reporters in catalytic reactions. Future challenges comprising the integration of other techniques, such as diffraction, scanning probe, or vibrational methods in single-molecule fluorescence spectroscopy are suggested.
荧光显微镜在时间和空间分辨率以及分子检测灵敏度方面的不断提高，为我们深入了解化学和生物催化剂的功能提供了独特的机会。由于单分子显微镜能够逐个计数周转事件，因此可以绘制固体多相催化剂中不同位点催化活性的分布图，或者研究酶或化学催化剂中单个位点随时间变化的活性波动。通过实验监测个体而非群体，可以成功阐明复杂行为的起源，例如动力学或失活过程中的行为。本文将根据单分子荧光显微镜在时间和空间分辨率方面的进展对催化测定的影响进行讨论。文中还举例说明了荧光报告分子在催化反应中的应用关键概念。同时提出了未来的挑战，包括将其他技术，如衍射、扫描探针或振动方法集成到单分子荧光光谱中。
