Chemistry Department, Brandeis University, MS 015, 415 South Street, Waltham, MA 02454, USA.
Curr Opin Chem Biol. 2009 Oct;13(4):436-42. doi: 10.1016/j.cbpa.2009.06.011.
Single molecule optical microscopy can directly monitor substrate turnover by individual enzymes revealing the underlying distribution of reaction rates and enzyme conformations. These techniques are particularly useful for assessing cooperativity in multi-subunit enzymes such as beta-galactosidase, and for directly monitoring how ligand and substrate binding alter dynamic equilibria. Recent investigations of HIV reverse transcriptase have reiterated the importance of single molecule microscopy for determining how proteins move on oligonucleotides and how ligands and inhibitors affect motion. Similar investigations of membrane active enzymes allow direct imaging of protein-membrane interactions. For a large variety of systems, single molecule enzymology provides unprecedented images of how enzymes interact with their substrates and the differences between individual enzymes in a population.
单分子光学显微镜可以直接监测单个酶的底物转化，揭示反应速率和酶构象的基础分布。这些技术对于评估β-半乳糖苷酶等多亚基酶的协同作用特别有用，并且可以直接监测配体和底物结合如何改变动态平衡。最近对 HIV 逆转录酶的研究再次强调了单分子显微镜在确定蛋白质如何在寡核苷酸上移动以及配体和抑制剂如何影响运动的重要性。对膜活性酶的类似研究允许直接成像蛋白质-膜相互作用。对于各种各样的系统，单分子酶学提供了前所未有的图像，展示了酶如何与它们的底物相互作用，以及在一个群体中个体酶之间的差异。
