Department of Chemical Engineering, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, 10617, Taiwan.
Amino Acids. 2010 Aug;39(3):821-9. doi: 10.1007/s00726-010-0536-0. Epub 2010 Mar 5.
This work examines the effects of L-arginine (L-Arg) on the aggregation and amyloid fibrillation of bovine serum albumin (BSA). We demonstrate that L-Arg dose-dependently reduces thioflavin T (ThT) fluorescence of BSA within the L-Arg concentration range used (0-1.4 M). However, as revealed by electron microscopy, size exclusion chromatography, and dynamic light scattering results, L-Arg does not prevent amyloid-like fibril formation by BSA. We conclude that L-Arg competes against ThT for binding sites on BSA amyloid-like fibrils, leading to biased results in ThT fluorescence measurements. Moreover, the use of ThT fluorescence assay to screen for potential inhibitors against amyloid fibrillation can give misleading results.
这项工作研究了 L-精氨酸（L-Arg）对牛血清白蛋白（BSA）聚集和淀粉样纤维形成的影响。我们证明，在使用的 L-Arg 浓度范围内（0-1.4 M），L-Arg 剂量依赖性地降低了 BSA 的硫黄素 T（ThT）荧光。然而，正如电子显微镜、尺寸排阻色谱和动态光散射结果所揭示的那样，L-Arg 并不能阻止 BSA 形成类似淀粉样的纤维。我们的结论是，L-Arg 与 ThT 竞争 BSA 类似淀粉样纤维上的结合位点，导致 ThT 荧光测量结果产生偏差。此外，使用 ThT 荧光测定法筛选潜在的抗淀粉样纤维形成抑制剂可能会产生误导性结果。
