Detection of protease activities by flash chronopotentiometry using a reversible polycation-sensitive polymeric membrane electrode.
Detection of protease activities by flash chronopotentiometry using a reversible polycation-sensitive polymeric membrane electrode.
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DOI：
10.1016/j.ab.2011.04.036
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发表时间：
2011-09-01
影响因子：
2.9
通讯作者：
Meyerhoff ME
中科院分区：
文献类型：
--
作者：
Gemene KL;Meyerhoff ME
一种新的电化学方法，称为闪光计时电位法（FCP），用于发展一种快速和灵敏的方法来检测蛋白酶的活性。在该方法中，在聚阳离子选择性聚合物膜上施加适当的电流脉冲，以诱导聚阳离子肽从样品相进入电极的有机膜的强通量。在该电流脉冲期间，连续监测细胞电位（EMF），并且其是多肽浓度的函数。施加的电流导致样品/膜界面处的多肽的局部耗尽，这在被称为过渡时间（τ）的特征时间处在观察到的计时电位图中产生剧烈的电位变化。对于给定的电流大小，τ的平方根与多肽的浓度成正比。蛋白酶将多肽切割成更小的片段，这些片段不能被有利地提取到传感器的膜中。因此，在蛋白水解过程中观察到过渡时间的减少。转变时间的变化程度可以与蛋白酶活性相关。为了证明这种方法，使用鱼精蛋白和合成的聚阳离子寡肽检测胰蛋白酶和α-胰凝乳蛋白酶的活性，所述聚阳离子寡肽具有被这些蛋白酶识别的特异性切割位点。
A novel electrochemical method, termed flash chronopotentiometry (FCP), is used to develop a rapid and sensitive method to detect protease activities. In this method, an appropriate current pulse is applied across a polycation-selective polymer membrane to induce a strong flux of the polycationic peptides from the sample phase into the organic membrane of the electrode. During this current pulse, the cell potential (EMF) is monitored continuously, and is a function of the polypeptide concentration. The imposed current causes a local depletion of the polypeptide at the sample/membrane interface, which yields a drastic potential change in the observed chronopotentiogram at a characteristic time, called the transition time (τ). For a given magnitude of current, the square root of τ is directly proportional to the concentration of the polypeptide. Proteases cleave polypeptides into smaller fragments that are not favorably extracted into the membrane of the sensor. Therefore, a decrease in the transition time is observed during the proteolysis process. The degree of change in the transition time can be correlated to protease activity. To demonstrate this approach, the activities of trypsin and α-chymotrypsin are detected using protamine and synthetic polycationic oligopeptides that possess specific cleavage sites that are recognized by these proteases.
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影响因子：
2.9
作者：
Lathia US;Ornatsky O;Baranov V;Nitz M
通讯作者：
Nitz M
影响因子：
2.9
作者：
DAVIE, EW;FUJIKAWA, K;KISIEL, W
通讯作者：
KISIEL, W
影响因子：
2.9
作者：
Lathia US;Ornatsky O;Baranov V;Nitz M
通讯作者：
Nitz M
影响因子：
15
作者：
Makarychev-Mikhailov, S;Shvarev, A;Bakker, E
通讯作者：
Bakker, E
影响因子：
4.3
作者：
Abuknesha, Ramadan A.;Jeganathan, Fiona;Price, Robert G.
通讯作者：
Price, Robert G.