Discerning Tyrosine Phosphorylation from Multiple Phosphorylations Using a Nanofluidic Logic Platform
Discerning Tyrosine Phosphorylation from Multiple Phosphorylations Using a Nanofluidic Logic Platform
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DOI：
10.1021/acs.analchem.1c03889
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发表时间：
2021-12-07
影响因子：
7.4
通讯作者：
Qing，Guangyan
中科院分区：
文献类型：
--
作者：
Xiong，Yuting;Li，Minmin;Qing，Guangyan
识别Tyr激酶催化的酪氨酸磷酸化（pTyr）对于揭示致癌机制和开发靶向抗癌药物至关重要。尽管有一些技术，但这一目标仍然具有挑战性，特别是当面临多种磷酸化事件的干扰时，包括丝氨酸（pSer）和苏氨酸磷酸化（pThr）。我们在这里描述了一种功能性聚合物修饰的人工离子纳米通道，它能够通过不同的离子电流变化敏感和选择性地识别磷酸酪氨酸（pY）肽。这种识别效应允许纳米通道在复杂的蛋白质消化条件下工作。此外，通过添加Ca 2+实现纳米流体逻辑功能显著提高了纳米通道对pY肽的选择性，因此可以通过在一锅法中同时监测相同或不同肽底物处的多位点磷酸化来区分Tyr激酶的pTyr和Ser/Thr激酶的pSer。该逻辑传感平台在区分Tyr激酶和Ser/Thr激酶以及在多靶点药物设计中评估多激酶活性方面具有潜力。
Discerning tyrosine phosphorylation (pTyr) catalyzed by Tyr kinase is central to the revelation of oncogenic mechanisms and the development of targeted anticancer drugs. Despite some techniques, this goal remains challenging, especially when faced with the interference of multiple phosphorylation events, including serine (pSer) and threonine phosphorylation (pThr). We describe here a functional polymer-modified artificial ion nanochannel, which enables the sensitive and selective recognition of phosphotyrosine (pY) peptide by the distinct ionic current change. Such a recognition effect allows for the nanochannel to work in a complex protein digest condition. Further, the implementation of nanofluidic logic functions with the addition of Ca2+dramatically improves the selectivity of the nanochannel to pY peptide and thus can discern pTyr by the Tyr kinase from pSer by the Ser/Thr kinase through simultaneously monitoring multisite phosphorylation at the same or different peptide substrates in one-pot. This logic sensing platform displays the potential in differentiating Tyr kinase and Ser/Thr kinase and assessing multi-kinase activities in multi-targeted drug design.