Meng Qinghe, Novak Adam, Rayhill Erin
SUNY Upstate Medical University, Syracuse, NY, USA.
Methods Mol Biol. 2025;2917:57-64. doi: 10.1007/978-1-0716-4478-2_5.
Zymography is a laboratory technique used to study the activity of enzymes, particularly proteases and glycosidases, in biological samples. It involves the separation of enzymatic proteins by gel electrophoresis under non-denaturing conditions, followed by their renaturation and incubation with specific substrates that mimic natural substrates. The resulting enzymatic activity leads to the degradation of the substrate within the gel, producing clear bands or zones of decreased staining that indicate the presence of active enzymes.There are several variants of zymography, including gelatin zymography, casein zymography, and substrate-specific zymography, each tailored to detect specific enzyme activities. Gelatin zymography, for example, is commonly used to detect protease activity against gelatin, while casein zymography employs casein as the substrate for a broader range of proteases.Zymography can be applied in various fields, including basic research, clinical diagnostics, and drug development. It provides valuable insights into enzyme function, substrate specificity, and regulation, and can serve as a biomarker for diseases associated with dysregulated enzyme activities.Overall, zymography is a versatile and widely used technique that offers qualitative information about enzyme activities and their role in biological processes. It contributes to our understanding of enzyme function and has potential applications in both research and clinical settings.
酶谱法是一种用于研究生物样品中酶(特别是蛋白酶和糖苷酶)活性的实验室技术。它包括在非变性条件下通过凝胶电泳分离酶蛋白,然后使其复性,并与模拟天然底物的特定底物一起孵育。产生的酶活性导致凝胶内底物的降解,产生清晰的条带或染色减少的区域,表明存在活性酶。酶谱法有几种变体,包括明胶酶谱法、酪蛋白酶谱法和底物特异性酶谱法,每种方法都针对检测特定的酶活性进行了优化。例如,明胶酶谱法通常用于检测针对明胶的蛋白酶活性,而酪蛋白酶谱法使用酪蛋白作为更广泛蛋白酶的底物。酶谱法可应用于各个领域,包括基础研究、临床诊断和药物开发。它为酶的功能、底物特异性和调节提供了有价值的见解,并可作为与酶活性失调相关疾病的生物标志物。总的来说,酶谱法是一种用途广泛且被广泛使用的技术,它提供了有关酶活性及其在生物过程中作用的定性信息。它有助于我们理解酶的功能,并在研究和临床环境中都有潜在的应用。