Kinetic parameters of Tau_244–372 fibrillization in the presence of different concentrations of full-length human PDI and its fragments as determined by ThT binding assays at 37°C.

Kinetic parameters, k, tm, and the lag time, were determined by fitting ThT fluorescence intensity versus time to Equation 1. The final concentration of human Tau fragment Tau244–372 was 10 µM. The buffer used was 10 mM HEPES buffer (pH 7.4) containing 1 mM DTT, 100 mM NaCl, 2.5 µM heparin and 20 µM ThT. Errors shown are standard errors of the mean. aKinetic parameters of Tau244–372 fibrillization in the presence of 100 g/l Ficoll 70 and different concentrations of full-length human PDI as determined by ThT binding assays at 37°C. ∼, Observed from the ThT fluorescence curves directly.通过将硫黄素T（Thioflavin T）的荧光强度随时间变化的曲线拟合至公式1，可得到动力学参数k、tm以及延滞时间。人源Tau片段Tau244–372的终浓度为10 µM。实验所用缓冲液为pH 7.4的10 mM羟乙基哌嗪乙磺酸（HEPES）缓冲液，体系中添加有1 mM二硫苏糖醇（DTT）、100 mM氯化钠（NaCl）、2.5 µM肝素（heparin）以及20 µM硫黄素T（Thioflavin T）。图中标注的误差为平均值的标准误。 a：于37℃下通过硫黄素T结合实验测定的，在100 g/l菲可70（Ficoll 70）与不同浓度全长人源蛋白二硫键异构酶（PDI）存在时，Tau244–372淀粉样纤维形成的动力学参数。 ~：直接由硫黄素T荧光曲线观测得到。