Department of Chemistry, East Carolina University, Greenville, North Carolina, United States.
Department of Chemistry, Davidson College, Davidson, North Carolina, United States.
Chembiochem. 2024 May 2;25(9):e202400007. doi: 10.1002/cbic.202400007. Epub 2024 Mar 28.
The actin cytoskeleton is a biosensor of cellular stress and a potential prognosticator of human disease. In particular, aberrant cytoskeletal structures such as stress granules formed in response to energetic and oxidative stress are closely linked to ageing, cancer, cardiovascular disease, and viral infection. Whether these cytoskeletal phenomena can be harnessed for the development of biosensors for cytoskeletal dysfunction and, by extension, disease progression, remains an open question. In this work, we describe the design and development of an optogenetic iteration of profilin, an actin monomer binding protein with critical functions in cytoskeletal dynamics. We demonstrate that this optically activated profilin ('OptoProfilin') can act as an optically triggered biosensor of applied cellular stress in select immortalized cell lines. Notably, OptoProfilin is a single component biosensor, likely increasing its utility for experimentalists. While a large body of preexisting work closely links profilin activity with cellular stress and neurodegenerative disease, this, to our knowledge, is the first example of profilin as an optogenetic biosensor of stress-induced changes in the cytoskeleton.
肌动蛋白细胞骨架是细胞应激的生物传感器，也是人类疾病的潜在预后因子。特别是，由于能量和氧化应激而形成的异常细胞骨架结构（例如应激颗粒）与衰老、癌症、心血管疾病和病毒感染密切相关。这些细胞骨架现象是否可以被利用来开发细胞骨架功能障碍的生物传感器，进而扩展到疾病进展，仍然是一个悬而未决的问题。在这项工作中，我们描述了一种原肌球蛋白的光遗传学迭代设计和开发，原肌球蛋白是一种肌动蛋白单体结合蛋白，在细胞骨架动力学中具有关键功能。我们证明，这种光激活的原肌球蛋白（“OptoProfilin”）可以作为一种光触发的生物传感器，用于检测选定的永生化细胞系中的细胞应激。值得注意的是，OptoProfilin 是一种单一成分的生物传感器，可能会增加其实验员的使用便利性。尽管大量现有的研究密切关联原肌球蛋白的活性与细胞应激和神经退行性疾病，但这是原肌球蛋白作为光遗传学生物传感器检测应激诱导的细胞骨架变化的第一个例子。
