González-Vera Juan A, Morris May C
Cell Cycle Biosensors & Inhibitors, Department of Amino Acids, Peptides and Proteins, Institute of Biomolecules Max Mousseron (IBMM) CNRS-UMR 5247, 15 Avenue Charles Flahault, Montpellier 34093, France.
Proteomes. 2015 Nov 4;3(4):369-410. doi: 10.3390/proteomes3040369.
Probing the dynamic activities of protein kinases in real-time in living cells constitutes a major challenge that requires specific and sensitive tools tailored to meet the particular demands associated with cellular imaging. The development of genetically-encoded and synthetic fluorescent biosensors has provided means of monitoring protein kinase activities in a non-invasive fashion in their native cellular environment with high spatial and temporal resolution. Here, we review existing technologies to probe different dynamic features of protein kinases and discuss limitations where new developments are required to implement more performant tools, in particular with respect to infrared and near-infrared fluorescent probes and strategies which enable improved signal-to-noise ratio and controlled activation of probes.
实时探测活细胞中蛋白激酶的动态活性是一项重大挑战，这需要有专门定制的特定且灵敏的工具，以满足与细胞成像相关的特殊需求。基因编码和合成荧光生物传感器的发展，提供了在其天然细胞环境中以非侵入方式、高空间和时间分辨率监测蛋白激酶活性的手段。在此，我们综述了用于探测蛋白激酶不同动态特征的现有技术，并讨论了需要新进展以实现性能更优工具的局限性，特别是关于红外和近红外荧光探针以及能够提高信噪比和实现探针可控激活的策略方面。
