Application of Gelatin Zymography in Nanotoxicity Research.
Application of Gelatin Zymography in Nanotoxicity Research.
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明胶酶谱法在纳米毒性研究中的应用。
DOI：
10.1007/978-1-4939-8916-4_8
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Mo,Yiqun
中科院分区：
文献类型：
--
作者：
Zhang,Yue;Wan,Rong;Zhang,Qunwei;Mo,Yiqun
明胶酶谱是一种相对简单、廉价和强大的技术，可以检测能够从各种生物来源中降解明胶的蛋白水解酶。基质金属蛋白酶家族中的两个成员，明胶酶A和明胶酶B，因其具有很强的明胶降解活性而被特别用于检测。基质金属蛋白酶-2和基质金属蛋白酶-9还能降解一些细胞外基质分子，包括IV、V和XI型胶原、层粘连蛋白和聚集素核心蛋白，因此它们在纳米毒性研究中具有重要意义。在这项技术中，蛋白质通过十二烷基硫酸钠(SDS)-聚丙烯酰胺凝胶电泳法(PAGE)分离，明胶酶被十二烷基硫酸钠(SDS)激活，消化凝胶中嵌入的明胶。考马斯亮蓝R-250染色后，降解区域在蓝色背景下可见清晰的条带。在这里，我们描述了明胶酶谱的详细程序。
Gelatin zymography is a relatively simple, inexpensive, and powerful technique to detect proteolytic enzymes capable of degrading gelatin from various biological sources. It has been used particularly to detect the two members of the matrix metalloproteinase family, MMP-2 (gelatinase A) and MMP-9 (gelatinase B), due to their potent gelatin-degrading activity. MMP-2 and MMP-9 are also able to degrade a number of extracellular matrix molecules including type IV, V, and XI collagens, laminin, and aggrecan core protein, thus making them important in the nanotoxicity research. In this technique, proteins are separated by sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE), and gelatinases, activated by SDS, digest gelatin embedded in the gel. After staining with Coomassie Brilliant Blue R-250, areas of degradation are visible as clear bands against a blue-stained background. Here, we describe the detailed procedure for gelatin zymography.
影响因子：
3.8
作者：
Wan, Rong;Mo, Yiqun;Zhang, Xing;Chien, Sufan;Tollerud, David J.;Zhang, Qunwei
通讯作者：
Zhang, Qunwei
通过将小的、确定的底物与凝胶基质化学连接，扩大酶谱法的用途。
影响因子：
7
作者：
V. Kaberdin;K. McDowall
通讯作者：
K. McDowall