Holistic Methods for the Analysis of cNMP Effects
Holistic Methods for the Analysis of cNMP Effects
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DOI：
10.1007/164_2015_42
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发表时间：
2017-01-01
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影响因子：
--
通讯作者：
Kostenis, Evi
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作者：
Grundmann, Manuel;Kostenis, Evi
环核苷酸单磷酸（cNMP）代表活细胞中的原型第二信使，并且充当具有广泛功能的分子开关。环磷酸腺苷和环磷酸鸟苷是描述得最好的环磷酸鸟苷;然而，越来越多的证据表明环磷酸鸟苷和环磷酸鸟苷在信号传导中也发挥着重要作用。尽管有研究努力，迄今为止，对这些非经典cNMP的生物学知之甚少，这至少部分是由于过去的方法学问题导致整个领域的挫折。直到最近，随着最先进技术的使用，才有可能恢复非经典的cNMP研究。虽然高灵敏度的检测方法揭示了哺乳动物细胞中cCMP和cUMP的相关水平，但有关生物效应物及其生理相互作用的知识仍然不完整。整体生物物理学读数无标记且以无偏的方式捕获细胞响应，其优点在于检测通过传统生物化学测定方法难以获得的细胞信号传导的隐藏方面。在本章中，我们介绍了动态质量再分布（DMR）技术，以探索细胞信号超越既定的受体控制机制。cCMP和cUMP的信号结构的共同和独特的功能进行了鉴定。此外，完整活细胞的整合反应揭示了迄今未公开的非典型cNMP的额外效应子。未来的研究将展示整体方法将如何整合到当代cNMP研究的方法库中。
Cyclic nucleotide monophosphates (cNMPs) typify the archetype second messenger in living cells and serve as molecular switches with broad functionality. cAMP and cGMP are the best-described cNMPs; however, there is a growing body of evidence indicating that also cCMP and cUMP play a substantial role in signal transduction. Despite research efforts, to date, relatively little is known about the biology of these noncanonical cNMPs, which is due, at least in part, to methodological issues in the past entailing setbacks of the entire field. Only recently, with the use of state-of-the-art techniques, it was possible to revive noncanonical cNMP research. While high-sensitive detection methods disclosed relevant levels of cCMP and cUMP in mammalian cells, knowledge about the biological effectors and their physiological interplay is still incomplete. Holistic biophysical readouts capture cell responses label-free and in an unbiased fashion with the advantage to detect concealed aspects of cell signaling that are arduous to access via traditional biochemical assay approaches. In this chapter, we introduce the dynamic mass redistribution (DMR) technology to explore cell signaling beyond established receptor-controlled mechanisms. Both common and distinctive features in the signaling structure of cCMP and cUMP were identified. Moreover, the integrated response of whole live cells revealed a hitherto undisclosed additional effector of the noncanonical cNMPs. Future studies will show how holistic methods will become integrated into the methodological arsenal of contemporary cNMP research.