Lipid-binding form is a key conformation to induce a programmed cell death initiated in tobacco BY-2 cells by a proteinaceous elicitor of cryptogein
Lipid-binding form is a key conformation to induce a programmed cell death initiated in tobacco BY-2 cells by a proteinaceous elicitor of cryptogein
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DOI：
10.1111/j.0031-9317.2004.00317.x
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发表时间：
2004-06-01
影响因子：
6.4
通讯作者：
Shioi, Y
中科院分区：
文献类型：
--
作者：
Hirasawa, K;Amano, T;Shioi, Y
隐地蛋白是隐地疫霉分泌的一种蛋白质类诱导子，它能诱导烟草细胞过敏性死亡。分析了两种隐地蛋白突变体，一种是新合成的突变体N93 A，其第93位Asn残基被替换为Ala，另一种是K13 V，其第13位Lys被替换为瓦尔。根据细胞外碱化、活性氧（AOS）的产生和死亡进展评估这些突变的影响。突变N93A导致细胞外碱化、AOS产生和细胞死亡进展的活性分别降低至71.0、74.6和24.5%。在K13 V突变的情况下，这些比率分别变为114、3.38和7.40%。突变体的脂质结合活性进行了分析，使用荧光脂质脱氢麦角甾醇。N93A和K13V的结果分别为野生型的38.3%和3.40%。这些结果表明，脂质结合形式是唯一的构象诱导AOS的生产和程序性细胞死亡的植物。
Cryptogein, a proteinaceous elicitor secreted by Phytophthora cryptogea, induces a remarkable hypersensitive cell death in tobacco cells. Two cryptogein mutants were analysed to characterize the induction mechanism of cell death; one was a newly synthesized mutant N93A whose 93rd Asn residue was changed to Ala, the other was K13V whose Lys at position 13 was replaced with Val. The effect of these mutations was evaluated in terms of extracellular alkalization, production of active oxygen species (AOS) and progression to death. The mutation N93A resulted in a reduction in activity to 71.0, 74.6 and 24.5% for original rates of extracellular alkalization, AOS production and cell death progression, respectively. In the case of the K13V mutation, these rates changed to 114, 3.38 and 7.40%, respectively. The lipid-binding activities of the mutants were analysed using fluorogenic lipid of dehydroergosterol. The results for N93A and K13V were 38.3 and 3.40% compared with the wild type, respectively. These findings indicate that the lipid-binding form was the only conformation to induce the production of AOS and programmed cell death in plants.