Kidrič Marjetka, Sabotič Jerica, Stevanović Branka
Department of Biotechnology, Jožef Stefan Institute, Jamova cesta 39, 1000 Ljubljana, Slovenia.
Department of Biotechnology, Jožef Stefan Institute, Jamova cesta 39, 1000 Ljubljana, Slovenia.
J Plant Physiol. 2014 Jul 15;171(12):998-1002. doi: 10.1016/j.jplph.2014.03.011. Epub 2014 Apr 16.
The unique response of desiccation-tolerant, or resurrection plants, to extreme drought is accompanied by major changes in the protein pool, raising the possibility of the involvement of proteases. We detected and characterized proteases present in their active state in leaf extracts of desiccated Ramonda serbica Panč., a resurrection plant from the Balkan Peninsula. Plants desiccated under laboratory conditions and maintained in anhydrobiosis for 4 and 14 months revived upon rehydration. Protease activities were determined spectrophotometrically in solution and by zymography on gels. Several endo- and aminopeptidases were detected and characterized by their pH profiles. Their enzyme class was determined using specific inhibitors. Those with higher activities were a serine endopeptidase active against Bz-Arg-pNA with a pH optimum around 9, and aminopeptidases optimally active at pHs from 7 to 9 against Leu-pNA, Met-pNA, Phe-pNA, Pro-pNA and Ala-pNA. The levels of their activities in leaf extracts from desiccated plants were significantly higher than those from rehydrated plants and from regularly watered plants, implying their involvement in the recovery of vegetative tissues from desiccation.
耐旱植物,即复苏植物,对极端干旱的独特反应伴随着蛋白质组的重大变化,这增加了蛋白酶参与其中的可能性。我们在巴尔干半岛的复苏植物塞尔维亚腊蒙达(Ramonda serbica Panč.)干燥叶片提取物中检测并鉴定了处于活性状态的蛋白酶。在实验室条件下干燥并在脱水状态下维持4个月和14个月的植物,复水后恢复生机。通过分光光度法在溶液中以及通过凝胶酶谱法测定蛋白酶活性。检测到几种内切酶和氨肽酶,并根据其pH谱进行了表征。使用特异性抑制剂确定它们的酶类别。活性较高的是一种对Bz-Arg-pNA有活性的丝氨酸内切酶,其最适pH约为9,以及在pH 7至9对Leu-pNA、Met-pNA、Phe-pNA、Pro-pNA和Ala-pNA最具活性的氨肽酶。干燥植物叶片提取物中的酶活性水平明显高于复水植物和正常浇水植物,这表明它们参与了营养组织从干燥状态的恢复。