College of Life Sciences, Shandong Provincial Key Laboratory of Animal Resistance Biology, Collaborative Innovation Center of Cell Biology in Universities of Shandong, Institute of Biomedical Sciences, Shandong Normal University, Jinan, China.
Thorac Cancer. 2020 Oct;11(10):2993-2997. doi: 10.1111/1759-7714.13643. Epub 2020 Sep 7.
Microtubules are the major cytoskeletal component in eukaryotes which are essential for a large spectrum of cellular activities. Monitoring the behavior of microtubules is helpful for a better understanding of the regulatory mechanism governing microtubule architecture and microtubule-based activities. Here, we characterized the binding capability of a modified heptapeptide from tau to both tubulin and microtubules and sought to develop it as a fluorescent peptide for monitoring microtubules.
To deliver the fluorescent peptide into the cells, a cell-penetrating peptide was conjugated to the modified heptapeptide from tau and synthesized. The affinity of the modified heptapeptide was determined by microscale thermophoresis. The microtubule labeling ability was determined by adding the peptide into the polymerized microtubule solutions or cultured HeLa cells.; RESULTS: Affinity determination revealed that the tau-derived peptide specifically bound to tubulin. In addition, the peptide was able to label polymerized microtubules in solution, although no obvious microtubule filaments were observed clearly in living cells, probably due to the inadequate affinity.
These results suggest that using a peptide-based strategy for imaging microtubules might be plausible and attempts to improve its affinity is warranted in the future.
微管是真核生物中主要的细胞骨架成分，对于多种细胞活动都是必不可少的。监测微管的行为有助于更好地理解调节微管结构和基于微管的活动的调控机制。在这里，我们描述了来自 tau 蛋白的改良七肽与微管蛋白和微管的结合能力，并试图将其开发为用于监测微管的荧光肽。
为了将荧光肽递送到细胞中，将穿膜肽与来自 tau 蛋白的改良七肽缀合并合成。通过微量热泳动测定来确定改良七肽的亲和力。通过将肽添加到聚合的微管溶液或培养的 HeLa 细胞中来确定微管标记能力。
亲和力测定表明，tau 衍生的肽特异性地与微管蛋白结合。此外，该肽能够在溶液中标记聚合的微管，尽管在活细胞中没有观察到明显的微管纤维，这可能是由于亲和力不足所致。
这些结果表明，使用基于肽的策略来成像微管可能是合理的，并且在未来有必要尝试提高其亲和力。
