Department of Biochemistry, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad, India.
Department of Functional and Organic Food, Institute of Human Nutrition Sciences, Warsaw University of Life Sciences, Warsaw, Poland.
Methods Mol Biol. 2024;2832:233-239. doi: 10.1007/978-1-0716-3973-3_17.
The regulation of proteolytic enzymes by protease inhibitors is crucial for maintaining the balance between protein synthesis and degradation, preventing uncontrolled proteolysis and fine-tuning cellular processes essential for optimal function and survival of the plants. It is known that the plant protease inhibitors activities are induced in defense of biotic as well as abiotic stresses. Thus, beyond their fundamental physiological functions, their involvement in stress responses, such as drought, cold, and salinity, is of equally significant. The X-ray film contact print method is an effective method for assessing various protease inhibitors exposed to stress conditions. In this approach, initially plant protease inhibitors will be separated using electrophoresis, and then the gel is treated with trypsin, which inhibits protease inhibitors. This gel when placed on X-ray film, the trypsin will digest the gelatin layer present on the film and the gelatinolytic activity stalled at the premises of protease inhibitors. This will provide the impression of the differentially expressed protease inhibitors in stress-treated plants.
蛋白酶抑制剂对蛋白水解酶的调控对于维持蛋白质合成与降解之间的平衡至关重要，可以防止失控的蛋白水解，并精细调节细胞过程，从而确保植物的最佳功能和生存。已知植物蛋白酶抑制剂的活性会在应对生物和非生物胁迫时被诱导。因此，除了它们的基本生理功能外，它们在应对干旱、寒冷和盐度等应激反应中的作用同样重要。X 射线胶片接触印片法是评估各种蛋白酶抑制剂在应激条件下的有效方法。在该方法中，首先使用电泳分离植物蛋白酶抑制剂，然后用胰蛋白酶处理凝胶，胰蛋白酶会抑制蛋白酶抑制剂。将该凝胶放在 X 射线胶片上时，胰蛋白酶会消化胶片上的明胶层，而明胶水解酶的活性则会在蛋白酶抑制剂存在的位置停止。这将在应激处理的植物中提供差异表达的蛋白酶抑制剂的印象。
