Department of Physics, University of Alberta, 11335 Saskatchewan Dr NW, Edmonton, Alberta, T6G 2M9, Canada.
Department of Electrical and Computer Engineering, University of Alberta, 9107-116 St, Edmonton, Alberta, T6G 2V4, Canada.
Small. 2021 Jan;17(1):e2003560. doi: 10.1002/smll.202003560. Epub 2020 Dec 9.
Tubulin is an electrostatically negative protein that forms cylindrical polymers termed microtubules, which are crucial for a variety of intracellular roles. Exploiting the electrostatic behavior of tubulin and microtubules within functional microfluidic and optoelectronic devices is limited due to the lack of understanding of tubulin behavior as a function of solvent composition. This work displays the tunability of tubulin surface charge using dimethyl sulfoxide (DMSO) for the first time. Increasing the DMSO volume fractions leads to the lowering of tubulin's negative surface charge, eventually causing it to become positive in solutions >80% DMSO. As determined by electrophoretic mobility measurements, this change in surface charge is directionally reversible, i.e., permitting control between -1.5 and + 0.2 cm (V s) . When usually negative microtubules are exposed to these conditions, the positively charged tubulin forms tubulin sheets and aggregates, as revealed by an electrophoretic transport assay. Fluorescence-based experiments also indicate that tubulin sheets and aggregates colocalize with negatively charged g-C N sheets while microtubules do not, further verifying the presence of a positive surface charge. This study illustrates that tubulin and its polymers, in addition to being mechanically robust, are also electrically tunable.
微管蛋白是一种带有负电荷的蛋白质,它形成圆柱形聚合物,称为微管,这对于各种细胞内的作用是至关重要的。由于缺乏对微管蛋白作为溶剂组成函数的行为的理解,利用功能微流控和光电设备中的微管蛋白和微管的静电行为受到限制。这项工作首次显示了使用二甲基亚砜 (DMSO) 来调节微管蛋白表面电荷的能力。随着 DMSO 体积分数的增加,微管蛋白的负表面电荷降低,最终导致其在 >80% DMSO 的溶液中变为正电荷。通过电泳迁移率测量确定,这种表面电荷的变化是定向可逆的,即允许在 -1.5 和 +0.2 cm (V s) 之间进行控制。当通常带负电荷的微管蛋白暴露于这些条件时,带正电荷的微管蛋白形成微管蛋白片和聚集体,如电泳输送测定所揭示的那样。基于荧光的实验也表明,微管蛋白片和聚集体与带负电荷的 g-C 3 N 片共定位,而微管则没有,进一步证实了存在正表面电荷。这项研究表明,除了具有机械强度外,微管蛋白及其聚合物也是可电调节的。