Department of Food Science and Nutrition, Faculty of Food and Agricultural Sciences, King Saud University, 2460, Riyadh 11451, Saudi Arabia.
Protein Research Chair, Department of Biochemistry, College of Science, King Saud University, Riyadh, Saudi Arabia.
Int J Biol Macromol. 2019 May 1;128:54-60. doi: 10.1016/j.ijbiomac.2019.01.097. Epub 2019 Jan 22.
The aim of this study was to investigate the effects of sodium dodecyl benzenesulfonate (SDBS) on hen egg white lysozyme (HEWL) fibrillogenesis at pH 7.4. HEWL fibrillogenesis in the presence of SDBS was characterized using several spectroscopic techniques (turbidity, light scattering, intrinsic fluorescence, ThT binding assay, ThT kinetics, far-UV CD, and transmission electron mmicroscopy). The turbidity and light scattering data revealed that SDBS induces aggregation in HEWL in dose-dependent manner. HEWL aggregation was seen at low SDBS concentrations (0.03 to 0.5 mM) but it was not observed at concentrations of SDBS at >0.6 mM. The ThT and TEM data clearly showed that the aggregates formed in the presence of SDBS had an amyloid-like morphology. From the CD analysis it was clear that low SDBS concentrations decreases the α-helical content while the β-sheet content increased. As the SDBS concentration further increased, the α-helical content increased again. The ThT kinetics analysis revealed that the HEWL monomer directly converted into the amyloid fibril without lag phase. All the spectroscopic and microscopic results support the finding that low concentrations of SDBS stimulate fibrillogenesis in HEWL, and that no fibrillogenesis occurs at higher SDBS concentrations.
本研究旨在探讨十二烷基苯磺酸钠(SDBS)对 pH 值为 7.4 时鸡卵清白蛋白溶菌酶(HEWL)纤维形成的影响。使用多种光谱技术(浊度、光散射、内源荧光、ThT 结合测定、ThT 动力学、远紫外 CD 和透射电子显微镜)对 SDBS 存在下的 HEWL 纤维形成进行了表征。浊度和光散射数据表明,SDBS 以剂量依赖的方式诱导 HEWL 聚集。在低浓度 SDBS(0.03 至 0.5 mM)下可以观察到 HEWL 聚集,但在浓度高于 0.6 mM 的 SDBS 下未观察到聚集。ThT 和 TEM 数据清楚地表明,在 SDBS 存在下形成的聚集体具有类淀粉样纤维形态。从 CD 分析可以清楚地看出,低浓度的 SDBS 降低了α-螺旋含量,而β-折叠含量增加。随着 SDBS 浓度进一步增加,α-螺旋含量再次增加。ThT 动力学分析表明,HEWL 单体直接转化为无延滞期的淀粉样纤维。所有光谱和显微镜结果都支持以下发现:低浓度的 SDBS 刺激 HEWL 的纤维形成,而在较高浓度的 SDBS 下则不会发生纤维形成。