Sanderson D G, Anderson L B, Gross E L
Biochim Biophys Acta. 1986 Dec 3;852(2-3):269-78. doi: 10.1016/0005-2728(86)90232-x.
Direct and mediated electrolysis of the protein plastocyanin at a gold filar electrode is described. The filar electrode used is of a unique design that allows potentiometric measurements, steady-state voltammetry and absorption spectrophotometry to be performed on a few microliters of solution containing 0.1-1.0 mM protein. As a result, we have determined the formal potential and diffusion coefficient of the blue copper protein, plastocyanin, to be 372 +/- 5 mV vs. normal hydrogen electrode and 8.9 X 10(-7) cm2 X s-1, respectively. The same value of the formal potential is obtained from a steady-state current experiment, an equilibrium spectrophotometric experiment, and a twin-electrode steady-state spectrophotometric experiment. The fact that the diffusion coefficient is measured under conditions of steady-state current, results in significant improvement in signal to background over techniques that monitor a transient current, while the potential is changing.
本文描述了在金丝电极上对蛋白质质体蓝素进行直接和介导电解的过程。所使用的丝状电极具有独特的设计，能够对几微升含有0.1 - 1.0 mM蛋白质的溶液进行电位测量、稳态伏安法和吸收分光光度法。结果，我们测定了蓝色铜蛋白质体蓝素的形式电位和扩散系数，相对于标准氢电极分别为372 ± 5 mV和8.9×10⁻⁷ cm²·s⁻¹。通过稳态电流实验、平衡分光光度实验和双电极稳态分光光度实验获得了相同的形式电位值。在稳态电流条件下测量扩散系数这一事实，相较于监测电位变化时瞬态电流的技术，显著提高了信号与背景的比值。
