Plant-derived human acetylcholinesterase-R provides protection from lethal organophosphate poisoning and its chronic aftermath
Plant-derived human acetylcholinesterase-R provides protection from lethal organophosphate poisoning and its chronic aftermath
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DOI：
10.1096/fj.07-8112com
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发表时间：
2007-09-01
期刊：
影响因子：
4.8
通讯作者：
Mor, Tsafrir S.
中科院分区：
文献类型：
--
作者：
Evron, Tama;Geyer, Brian C.;Mor, Tsafrir S.
具有治疗价值的蛋白质在其自然环境中通常是稀有和/或不稳定的，需要在异源系统中生产解决方案。一个相关的例子是应激诱导的、通常罕见的、自然不稳定的人乙酰胆碱酯酶变异体AChE-R。AChE-R与突触AChE-S突变体共享其活性部位，后者是有毒的有机磷抗胆碱酯酶杀虫剂如对硫磷代谢物对氧磷的靶标。有机磷中毒下固有的AChE-R过量产生既提供了短期保护(作为生物清道夫)，也提供了长期神经肌肉损伤(作为调节)。在这里，我们报道了从烟草植物中提取的保留内质网的人AChE-RER的纯化、鉴定和检测。纯化到均一的AChE-RER表现出难以区分的生化特性，对氧磷脂的IC50=10(-7)M，类似于哺乳动物细胞培养来源的AChE。体内滴定显示，静脉注射AChE-RER对FVB/N雄性小鼠具有剂量依赖的保护作用，在接近摩尔等量的情况下，可完全消除短期临床症状。在暴露后10天，AChE-R预防明显限制了暴露后小鼠血浆AChE-R水平的增加，同时最大限度地减少了有机磷诱导的神经肌肉接头的形态异常。我们的发现表明，植物生产的AChE-RER是一种双峰剂，提供了对有机磷中毒的短期和长期保护。-Evron，T.，Geyer，B.C.，Cherni，I.，Muralidharan，M.，Kilbourne，J.，Fletcher，S.P.，Soreq，H.，Mor，T.S.植物来源的人乙酰胆碱酯酶-R提供了对致命性有机磷中毒及其慢性后果的保护。
Therapeutically valuable proteins are often rare and/or unstable in their natural context, calling for production solutions in heterologous systems. A relevant example is that of the stress-induced, normally rare, and naturally unstable "read-through" human acetylcholinesterase variant, AChE-R. AChE-R shares its active site with the synaptic AChE-S variant, which is the target of poisonous organophosphate anticholinesterase insecticides such as the parathion metabolite paraoxon. Inherent AChE-R overproduction under organophosphate intoxication confers both short-term protection ( as a bioscavenger) and long-term neuromuscular damages ( as a regulator). Here we report the purification, characterization, and testing of human, endoplasmic reticulumretained AChE-RER produced from plant-optimized cDNA in Nicotiana benthamiana plants. AChE-RER purified to homogeneity showed indistinguishable biochemical properties, with IC50 = 10(-7) M for the organophosphate paraoxon, similar to mammalian cell culture-derived AChE. In vivo titration showed dose-dependent protection by intravenously injected AChE-RER of FVB/N male mice challenged with a lethal dose of paraoxon, with complete elimination of short-term clinical symptoms at near molar equivalence. By 10 days postexposure, AChE-R prophylaxis markedly limited postexposure increases in plasma murine AChE-R levels while minimizing the organophosphate-induced neuromuscular junction dismorphology. Our findings present plant-produced AChE-RER as a bimodal agent, conferring both short- and long- term protection from organophosphate intoxication.-Evron, T., Geyer, B. C., Cherni, I., Muralidharan, M., Kilbourne, J., Fletcher, S. P., Soreq, H., Mor, T. S. Plant- derived human acetylcholinesterase- R provides protection from lethal organophosphate poisoning and its chronic aftermath.