Institute of Biosciences and Technology, Texas A&M University, Houston, Texas 77030, United States.
ORBIT Platform, The University of Texas MD Anderson Cancer Center, Houston, Texas 77054, United States.
ACS Sens. 2024 Sep 27;9(9):4758-4766. doi: 10.1021/acssensors.4c01167. Epub 2024 Aug 15.
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.
微管（MT）动力学受到微管相关蛋白（MAPs）和微管蛋白各种翻译后修饰（PTMs）的严格调控。在这里，我们引入寡聚 MT 和寡聚 TIP，它们是为实时可视化和操纵活细胞内 MT 行为而设计的基因编码寡聚 MT 结合物。寡聚 MT 可作为标记 MT 细胞骨架的可靠标记物，而寡聚 TIP 则可用于实时监测生长中的 MT 正极。这些工程化的 MT 结合物已成功用于标记 MT 网络、监测细胞分裂、跟踪 MT 正极末端，并评估微管乙酰化对 MT 稳定性的影响，达到单细胞水平。此外，寡聚 MT 和寡聚 TIP 可以被重新用作定量评估药物作用和报告酶活性的生物传感器。总之，这些工程化的 MT 结合物有望推进 MT 生物学的机制剖析，并在基于细胞的高通量药物发现工作中有转化应用。
