A self-referencing biosensor for real-time monitoring of physiological ATP transport in plant systems.
A self-referencing biosensor for real-time monitoring of physiological ATP transport in plant systems.
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DOI：
10.1016/j.bios.2015.05.027
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发表时间：
2015-12
影响因子：
12.6
通讯作者：
D. Vanegas;G. Clark;A. Cannon;S. Roux;P. Chaturvedi;E. McLamore
中科院分区：
文献类型：
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作者：
D. Vanegas;G. Clark;A. Cannon;S. Roux;P. Chaturvedi;E. McLamore
本研究的目的是开发一种自参考的电化学生物传感器，用于直接测量活细胞/生物进入细胞外基质的ATP通量。该生物传感器的工作机理是基于甘油-3-磷酸酶和甘油-3-磷酸酶的活性。在石墨烯修饰的铂电极上，采用以蛋白质为模板的硅溶胶凝胶包裹技术，制备了层状双酶纳米复合材料。该传感器具有良好的电化学性能，其灵敏度为2.4±1.8nA/µM，响应时间为2 0±13 nA/S，检测下限为1.3±0.7nM。自参比生物传感器通过(I)萌发蕨类孢子和(Ii)生长玉米来测量外源ATP的外流。树根。这篇手稿展示了首次开发的非侵入性三磷酸腺苷微型生物传感器，用于直接测量eATP在活组织中的转运。在这项工作之前，eATP的测定还不能记录生理水平(NM和亚NM)的ATP的瞬时运动。该方法准确地测量了植物细胞附近的[eATP]通量。尽管这些概念验证实验侧重于植物组织，但这里开发的技术适用于任何活组织，在这些组织中，纳米分子浓度的ATP在信号和发育中发挥关键作用。这一工具对于进行假说驱动的生命科学研究将是非常有价值的，该研究旨在了解ATP在细胞外环境中的作用。
The objective of this study was to develop a self-referencing electrochemical biosensor for the direct measurement of ATP flux into the extracellular matrix by living cells/organisms. The working mechanism of the developed biosensor is based on the activity of glycerol kinase and glycerol-3-phosphate oxidase. A stratified bi-enzyme nanocomposite was created using a protein-templated silica sol gel encapsulation technique on top of graphene-modified platinum electrodes. The biosensor exhibited excellent electrochemical performance with a sensitivity of 2.4±1.8 nA/µM, a response time of 20±13 s and a lower detection limit of 1.3±0.7 nM. The self-referencing biosensor was used to measure exogenous ATP efflux by (i) germinatingCeratopterisspores and (ii) growingZeamaysL. roots. This manuscript demonstrates the first development of a non-invasive ATP micro-biosensor for the direct measurement of eATP transport in living tissues. Before this work, assays of eATP have not been able to record the temporally transient movement of ATP at physiological levels (nM and sub-nM). The method demonstrated here accurately measured [eATP] flux in the immediate vicinity of plant cells. Although these proof of concept experiments focus on plant tissues, the technique developed herein is applicable to any living tissue, where nanomolar concentrations of ATP play a critical role in signaling and development. This tool will be invaluable for conducting hypothesis-driven life science research aimed at understanding the role of ATP in the extracellular environment.