细胞骨架对于广泛的细胞和组织水平过程的空间和时间组织至关重要。如扩散,信令,货物运输,迁移,形态发生,和神经元发育。细胞骨架研究旨在通过成像研究这些过程,或者通过本地操纵,具有高时空分辨率的细胞骨架蛋白的动力学和组织:与光学方法的能力相匹配。迄今为止,没有光响应微管稳定工具可以将实用的高精度试剂所需的所有功能结合在一起:低效力和生物化学稳定的非照明状态;然后是有效的,快速,和清洁的光响应,产生高效力的光照状态;加上良好的溶解度在合适的工作浓度;和有效的合成访问。我们现在介绍CouEpo,一种结合了这些需求的光老化埃坡霉素微管稳定试剂。在紫色/蓝色照射下,其效力增加约100倍,达到低纳摩尔值,允许对活细胞中的微管动力学进行有效的光控,甚至微管结构和细胞动力学中细胞不对称性的产生。因此,CouEpo是一种高性能的工具化合物,可以支持对许多微管相关过程的高精度研究,从生物物理学到运输,细胞运动性,和神经元生理学。
The cytoskeleton is essential for spatial and temporal organisation of a wide range of cellular and tissue-level processes, such as proliferation, signalling, cargo transport, migration, morphogenesis, and neuronal development. Cytoskeleton research aims to study these processes by imaging, or by locally manipulating, the dynamics and organisation of cytoskeletal proteins with high spatiotemporal resolution: which matches the capabilities of optical methods. To date, no photoresponsive microtubule-stabilising tool has united all the features needed for a practical high-precision reagent: a low potency and biochemically stable non-illuminated state; then an efficient, rapid, and clean photoresponse that generates a high potency illuminated state; plus good solubility at suitable working concentrations; and efficient synthetic access. We now present CouEpo, a photocaged epothilone microtubule-stabilising reagent that combines these needs. Its potency increases approximately 100-fold upon violet/blue irradiation to reach low-nanomolar values, allowing efficient photocontrol of microtubule dynamics in live cells, and even the generation of cellular asymmetries in microtubule architecture and cell dynamics. CouEpo is thus a high-performance tool compound that can support high-precision research into many microtubule-associated processes, from biophysics to transport, cell motility, and neuronal physiology.