In situ demonstration and quantitative analysis of the intrinsic properties of glycoside hydrolases
In situ demonstration and quantitative analysis of the intrinsic properties of glycoside hydrolases
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DOI：
10.1002/elps.201100333
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发表时间：
2012-01
期刊：
影响因子：
2.9
通讯作者：
Xiaomei Zhang;Ning Liu;Fan Yang;Jinghua Li;Lushan Wang;Guanjun Chen;P. Gao
中科院分区：
文献类型：
--
作者：
Xiaomei Zhang;Ning Liu;Fan Yang;Jinghua Li;Lushan Wang;Guanjun Chen;P. Gao
基于数字图像分析技术和一系列非变性电泳优化，建立了一种同时检测糖苷水解酶酶系中各活性组分内在性质的直接新方法。其关键技术是通过电泳分离后相应条带的灰度值变化来定量测定凝胶上底物（或产物）的浓度变化。以这种方式，每个糖苷水解酶组分的催化特性被证明在原位和容易确定后，浸泡在一系列的溶液中含有底物或其衍生物的凝胶。该方法具有通量高、灵敏度高、操作简便等优点，可用于研究糖苷水解酶的天然多样性和多酶表达系统的时空调控。因此，研究糖苷水解酶的功能蛋白质组学是一条有效的途径。
Based on digital image analysis techniques and a series of optimizations in native electrophoresis, a new direct method to simultaneously detect the intrinsic properties of each active component in the enzymatic system of glycoside hydrolase was established. The key technique is that the concentration changes of substrate (or product) on the gel can be determined quantitatively by the gray value changes of the corresponding band after electrophoretic separation. In this manner, the catalytic characteristics of each glycoside hydrolase component were demonstrated in situ and were easily determined after immersing the gel in a series of solutions containing substrates or their derivatives. Because of its high throughput, great sensitivity, and convenient operation, this method can be used to demonstrate the natural diversity of glycoside hydrolases and to study spatial and temporal regulation in multienzyme expression systems. Thus, it is an effective approach to study the functional proteomics of glycoside hydrolases.