Purification, characterization and identification of a senescence related serine protease in dark-induced senescent wheat leaves

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• 作者：Renxian Wang ; Shaowei Liu ; Jin Wang ; Qiang Dong ; Langlai Xu ; Qi Rui
• 关键词：EP ; endopeptidase ; PAGE ; polyacrylamide gel electrophoresis ; SDS ; sodium dodecylsulfate ; DMSO ; dimethyl sulfoxide ; DTT ; dithiothreitol ; AEBSF ; 4-(2-aminoethyl) benzene sulfonyl fluoride hydrochloride ; PMSF ; phenylmethanesulfonyl fluoride ; EGTA ; ethylene g
• 刊名：Phytochemistry
• 出版年：2013
• 出版时间：November, 2013
• 年：2013
• 卷：95
• 期：Complete
• 页码：118-126
• 全文大小：2110 K

文摘

Senescence-related proteases play important roles in leaf senescence by regulating protein degradation and nutrient recycling. A 98.9 kDa senescence-related protease EP3 in wheat leaves was purified by ammonium sulfate precipitation, Q-Sepharose fast flow anion exchange chromatography and gel slicing after gel electrophoresis. Due to its relatively high thermal stability, its protease activity did not decrease after incubation at 40 ¡ãC for 1-h. EP3 protease was suggested to be a metal-dependent serine protease, because its activity was inhibited by serine protease inhibitors PMSF and AEBSF and metal related protease inhibitor EGTA. It was identified as a subtilisin-like serine protease of the S8A family based on data from both mass spectrometry and the cloned cDNA sequence. Therefore, these data suggest that a serine protease of the S8A subfamily with specific biochemical properties is involved in senescence-associated protein degradation.