Davis J J, Burgess H, Zauner G, Kuznetsova S, Salverda J, Aartsma T, Canters G W
Central Research Laboratory, Mansfield Road, Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3QR, United Kingdom.
J Phys Chem B. 2006 Oct 19;110(41):20649-54. doi: 10.1021/jp0630525.
Generation of functionally active biomolecular monolayers is important in both analytical science and biophysical analyses. Our ability to monitor the redox-active state of immobilized proteins or enzymes at a molecular level, from which stochastic and surface-induced variations would be apparent, is impeded by comparatively slow electron-transfer kinetics and associated signal:noise difficulties. We demonstrate herein that by covalently tethering an appropriate dye to the copper protein azurin a highly oxidation-state-sensitive FRET process can be established which enables redox switching to be optically monitored at protein levels down to the zeptomolar limit. The surface-potential-induced cycling of emission enables the redox potential of clusters of a few hundred molecules to be determined.
功能活性生物分子单层的生成在分析科学和生物物理分析中都很重要。我们在分子水平上监测固定化蛋白质或酶的氧化还原活性状态的能力受到相对较慢的电子转移动力学以及相关的信号噪声问题的阻碍，在这种情况下，随机和表面诱导的变化会很明显。我们在此证明，通过将合适的染料共价连接到铜蛋白天青蛋白上，可以建立一种对高氧化态敏感的荧光共振能量转移（FRET）过程，该过程能够在低至zeptomolar极限的蛋白质水平上对氧化还原开关进行光学监测。表面电位诱导的发射循环能够确定几百个分子簇的氧化还原电位。
