Electrochemical proteolytic beacon for detection of matrix metalloproteinase activities
Electrochemical proteolytic beacon for detection of matrix metalloproteinase activities
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DOI：
10.1021/ja0626638
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发表时间：
2006-09-27
影响因子：
15
通讯作者：
Lin, Yuehe
中科院分区：
文献类型：
--
作者：
Liu, Guodong;Wang, Jun;Lin, Yuehe
本通讯描述了一种利用二茂铁报告物标记的肽底物检测基质金属蛋白酶-7活性的新方法。该底物可作为该金属蛋白酶的选择性“电化学蛋白水解信标”（EPB）。EPB固定在金电极表面，以实现对未裂解和裂解事件的“开-关”电化学信号能力。通过二茂铁氧化还原电流的下降速率测定了MMP-7对EPB的有效和选择性切割。直接将肽裂解事件对应的信号转导为电子信号，从而为检测MMP活性提供了一种简单、敏感的途径。新方法允许在低至3.4 pM的浓度下鉴定MMP-7的活性。这个概念可以扩展到设计一个多肽底物，用不同的电活性报告物标记，用于分析多种MMPs的活性。
This communication describes a novel method for detecting matrix metalloproteinase-7 activity using a peptide substrate labeled with a ferrocene reporter. The substrate serves as a selective “electrochemical proteolytic beacon” (EPB) for this metalloproteinase. The EPB is immobilized on a gold electrode surface to enable “on−off” electrochemical signaling capability for uncleaved and cleaved events. The EPB is efficiently and selectively cleaved by MMP-7 as measured by the rate of decrease in redox current of ferrocene. Direct transduction of a signal corresponding to peptide cleavage events into an electronic signal thus provides a simple, sensitive route for detecting the MMP activity. The new method allows for identification of the activity of MMP-7 in concentrations as low as 3.4 pM. The concept can be extended to design a multiple peptide substrate labeled with different electroactive reporters for assaying multiple MMPs activities.