Institute of Medical Genetics, Center for Pathobiochemistry and Genetics, Medical University of Vienna, Vienna, Austria.
Vienna Doctoral School of Pharmaceutical, Nutritional and Sport Sciences (PhaNuSpo), University of Vienna, Vienna, Austria.
Protein Sci. 2024 Oct;33(10):e5170. doi: 10.1002/pro.5170.
The intrinsically disordered protein MeCP2 is a global transcriptional regulator encoded by the MECP2 gene. Although the structured domains of MeCP2 have been the subject of multiple studies, its unstructured regions have not been that extensively characterized. In this work, we show that MeCP2 possesses properties akin to those of supercharged proteins. By utilizing its unstructured portions, MeCP2 can successfully transduce across cell membranes and localize to heterochromatic foci in the nuclei, displaying uptake levels a third lower than a MeCP2 construct fused to the cell-penetrating peptide TAT. MeCP2 uptake can further be enhanced by the addition of compounds that promote endosomal escape following cellular trafficking by means of macropinocytosis. Using a combination of in silico prediction algorithms and live-cell imaging experiments, we mapped the sequence in MeCP2 responsible for its cellular incorporation, which bears a striking resemblance to TAT itself. Transduced MeCP2 was shown to interact with HDAC3. These findings provide valuable insight into the properties of MeCP2 and may be beneficial for devising future protein-based treatment strategies.
天然无序蛋白 MeCP2 是由 MECP2 基因编码的一种全局转录调控因子。虽然 MeCP2 的结构域已经经过多次研究,但它的无结构区域尚未得到广泛的描述。在这项工作中,我们表明 MeCP2 具有类似于超荷电蛋白的特性。通过利用其无结构部分,MeCP2 可以成功地跨细胞膜转导,并在核内定位到异染色质焦点,其摄取水平比融合了穿膜肽 TAT 的 MeCP2 构建体低三分之一。通过添加促进细胞吞噬作用后内体逃逸的化合物,可以进一步增强 MeCP2 的摄取。我们使用计算机预测算法和活细胞成像实验相结合的方法,绘制了 MeCP2 中负责其细胞摄取的序列图谱,该序列与 TAT 本身惊人地相似。转导的 MeCP2 被证明与 HDAC3 相互作用。这些发现为 MeCP2 的特性提供了有价值的见解,并可能有助于设计未来基于蛋白质的治疗策略。