Genetically Encoded Microtubule Binders for Single-Cell Interrogation of Cytoskeleton Dynamics and Protein Activity,ACS Sensors

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Genetically Encoded Microtubule Binders for Single-Cell Interrogation of Cytoskeleton Dynamics and Protein Activity
ACS Sensors ( IF 8.2 ) Pub Date : 2024-08-15 , DOI: 10.1021/acssensors.4c01167
Joseph Zhou ₁ , Xiaoxuan Liu ₁ , Dekai Zhang ₁ , Guolin Ma ₂

Affiliation

Microtubule (MT) dynamics is tightly regulated by microtubule-associated proteins (MAPs) and various post-translational modifications (PTMs) of tubulin. Here, we introduce OligoMT and OligoTIP as genetically encoded oligomeric MT binders designed for real-time visualization and manipulation of MT behaviors within living cells. OligoMT acts as a reliable marker to label the MT cytoskeleton, while OligoTIP allows for live monitoring of the growing MT plus-ends. These engineered MT binders have been successfully utilized to label the MT network, monitor cell division, track MT plus-ends, and assess the effect of tubulin acetylation on the MT stability at the single-cell level. Moreover, OligoMT and OligoTIP can be repurposed as biosensors for quantitative assessment of drug actions and for reporting enzymatic activity. Overall, these engineered MT binders hold promise for advancing the mechanistic dissection of MT biology and have translational applications in cell-based high-throughput drug discovery efforts.

中文翻译：

用于单细胞研究细胞骨架动力学和蛋白质活性的基因编码微管结合物

微管 (MT) 动力学受到微管相关蛋白 (MAP) 和微管蛋白的各种翻译后修饰 (PTM) 的严格调控。在这里，我们介绍 OligoMT 和 OligoTIP 作为基因编码的寡聚 MT 结合剂，旨在实时可视化和操纵活细胞内的 MT 行为。 OligoMT 是标记 MT 细胞骨架的可靠标记，而 OligoTIP 可以实时监测不断生长的 MT 末端。这些工程化的 MT 结合剂已成功用于标记 MT 网络、监测细胞分裂、跟踪 MT 加端，并评估微管蛋白乙酰化对单细胞水平 MT 稳定性的影响。此外，OligoMT 和 OligoTIP 可以重新用作生物传感器，用于定量评估药物作用和报告酶活性。总体而言，这些工程化的 MT 结合剂有望推进 MT 生物学的机械解剖，并在基于细胞的高通量药物发现工作中具有转化应用。

更新日期：2024-08-15

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