Hatsugai Noriyuki, Hara-Nishimura Ikuko
Department of Plant and Microbial Biology, Microbial and Plant Genomics Institute, University of Minnesota, St. Paul, MN, USA.
Faculty of Science and Engineering, Konan University, Kobe, Japan.
Methods Mol Biol. 2018;1743:163-171. doi: 10.1007/978-1-4939-7668-3_15.
Caspase-like activities are essential to regulate programed cell death in plants. Although no caspase orthologous enzymes with aspartic acid specificity have been identified in plants, vacuolar processing enzyme (VPE) exhibits a caspase-1-like activity. In this chapter, we introduce two methods for the measurement of the caspase-1-like/VPE activity. These methods are based on the cleavage of caspase-1 specific synthetic substrates and on monitoring the active forms of VPE using a biotinylated-inhibitor blot analysis. Both methods are also adaptable to other plant caspase-like activities.
类半胱天冬酶活性对于调节植物程序性细胞死亡至关重要。尽管在植物中尚未鉴定出具有天冬氨酸特异性的半胱天冬酶直系同源酶,但液泡加工酶(VPE)表现出类半胱天冬酶-1活性。在本章中,我们介绍了两种用于测量类半胱天冬酶-1/VPE活性的方法。这些方法基于半胱天冬酶-1特异性合成底物的切割以及使用生物素化抑制剂印迹分析监测VPE的活性形式。这两种方法也适用于其他植物类半胱天冬酶样活性。