Seeing the forest for the trees: fluorescence studies of single enzymes in the context of ensemble experiments.
Seeing the forest for the trees: fluorescence studies of single enzymes in the context of ensemble experiments.
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DOI：
10.1039/c0cp02412k
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发表时间：
2011-01
期刊：
影响因子：
--
通讯作者：
Yan-Wen Tan;Haw Yang
中科院分区：
文献类型：
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作者：
Yan-Wen Tan;Haw Yang
酶是一种非凡的分子机器，它在温和的生物条件下以令人难以置信的精度和效率使许多困难的生化反应成为可能。然而，我们对酶的工作原理的了解还没有达到可以轻易地从酶的结构推断出其机理和催化速率的程度。解决与酶的三维结构及其功能相关的动力学问题已被确定为一个关键问题。尽管实施起来仍然具有挑战性，但单分子技术已经成为研究酶的最有用的方法之一。我们回顾了使用单分子荧光方法研究的酶，但将它们置于其他对大量样品进行的补充实验工作的结果的背景下。这篇综述主要涵盖了三种酶系统——黄酮类酶、去氢叶酸还原酶和腺苷酸激酶——并在适当的地方提到了其他酶。当将单分子实验与针对同一科学问题的其他方法放在一起讨论时，它们的弱点、优势和独特贡献就变得清晰起来。
Enzymes are remarkable molecular machines that make many difficult biochemical reactions possible under mild biological conditions with incredible precision and efficiency. Our understanding of the working principles of enzymes, however, has not reached the level where one can readily deduce the mechanism and the catalytic rates from an enzyme's structure. Resolving the dynamics that relate the three-dimensional structure of an enzyme to its function has been identified as a key issue. While still challenging to implement, single-molecule techniques have emerged as one of the most useful methods for studying enzymes. We review enzymes studied using single-molecule fluorescent methods but placing them in the context of results from other complementary experimental work done on bulk samples. This review primarily covers three enzyme systems--flavoenzymes, dehydrofolate reductase, and adenylate kinase--with additional enzymes mentioned where appropriate. When the single-molecule experiments are discussed together with other methods aiming at the same scientific question, the weakness, strength, and unique contributions become clear.