Toth Marta, Sohail Anjum, Fridman Rafael
Department of Pathology, School of Medicine, Proteases and Cancer Program, Karmanos Cancer Institute, Wayne State University, Detroit, MI, USA.
Methods Mol Biol. 2012;878:121-35. doi: 10.1007/978-1-61779-854-2_8.
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.
明胶酶谱法是一种简单却强大的方法，用于检测能够降解来自各种生物来源明胶的蛋白水解酶。由于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）具有强大的明胶降解活性，该方法对于评估它们尤为有用。这种基于聚丙烯酰胺凝胶电泳的方法能够可靠地评估培养细胞、组织和生物体液中明胶酶的类型、相对含量以及激活状态（与活性酶形式相比的潜伏状态）。该方法可用于研究在实验系统中调节明胶酶表达和调节酶原激活的因素。该系统可提供有关人类癌症组织中明胶酶表达和激活模式的信息，以及这与癌症进展的关系。对明胶酶谱法获得的数据进行解读需要透彻理解该技术的原理和陷阱；在评估酶水平和活性明胶酶种类的存在时，这一点尤为重要。如果使用得当，明胶酶谱法是研究生物系统中明胶酶的优秀工具。
