Assessment of gelatinases (MMP-2 and MMP-9) by gelatin zymography.
Assessment of gelatinases (MMP-2 and MMP-9) by gelatin zymography.
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DOI：
10.1007/978-1-61779-854-2_8
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Fridman, Rafael
中科院分区：
文献类型：
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作者：
Toth, Marta;Sohail, Anjum;Fridman, Rafael
明胶酶谱法是一种简单而强大的方法，用于检测能够降解各种生物来源的明胶的蛋白水解酶。由于其有效的明胶降解活性，它对于评估基质金属蛋白酶家族的两个关键成员 MMP-2（明胶酶 A）和 MMP-9（明胶酶 B）特别有用。这种基于聚丙烯酰胺凝胶电泳的方法可以对培养细胞、组织和生物液体中的明胶酶类型、相对量和激活状态（与活性酶形式相比是潜在的）提供可靠的评估。该方法可用于研究实验系统中调节明胶酶表达和调节酶原激活的因素。该系统提供有关人类癌症组织中明胶酶表达和激活模式及其与癌症进展的关系的信息。解释明胶酶谱分析中获得的数据需要彻底了解该技术的原理和缺陷；在评估酶水平和活性明胶酶种类的存在时，这一点尤其重要。如果使用得当，明胶酶谱法是研究生物系统中明胶酶的绝佳工具。
Gelatin zymography is a simple yet powerful method to detect proteolytic enzymes capable of degrading gelatin from various biological sources. It is particularly useful for the assessment of two key members of the matrix metalloproteinase family, MMP-2 (gelatinase A) and MMP-9 (gelatinase B), due to their potent gelatin-degrading activity. This polyacrylamide gel electrophoresis-based method can provide a reliable assessment of the type of gelatinase, relative amount, and activation status (latent, compared with active enzyme forms) in cultured cells, tissues, and biological fluids. The method can be used to investigate factors that regulate gelatinase expression and modulate zymogen activation in experimental systems. The system provides information on the pattern of gelatinase expression and activation in human cancer tissues and how this relates to cancer progression. Interpretation of the data obtained in gelatin zymography requires a thorough understanding of the principles and pitfalls of the technique; this is particularly important when evaluating enzyme levels and the presence of active gelatinase species. If properly used, gelatin zymography is an excellent tool for the study of gelatinases in biological systems.