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Author:
Date:
2021-01-05
[Abstract] Isoprenoids represent the largest class of metabolites with amazing diversities in structure and function. They are involved in protecting plants against pathogens or herbivores or involved in attracting pollinators. Isoprenoids are derived from geranyl diphosphate (GPP; C10), farnesyl diphosphate (FPP; C15), geranylgeranyl diphosphate (GGPP; C20), and geranylfarnesyl diphosphate (GFPP; C25) that are in turn formed by sequential condensations of isopentenyl diphosphate (IPP; C5) with an allylic acceptor such as dimethylallyl diphosphate (DMAPP; C5), GPP, FPP, or GGPP in a reaction catalyzed by isoprenyl diphosphate synthases (IDSs). IDS enzyme assay for determination of prenyl diphosphate products is generally performed ...
[摘要] [Abstrac吨]类异戊二烯代表最大的一类代谢物在结构和功能惊人多样性。它们参与保护植物免受病原体或草食动物侵害，或参与吸引传粉媒介。（;çGPP类异戊二烯是从牻牛儿基二磷酸衍生的10 ），法呢基二磷酸（FPP;Ç 15 ），香叶基香叶基二磷酸（GGPP;Ç 20 ），和geranylfarnesyl二磷酸（GFPP; C ^ 25 ），它们又通过的顺序缩合形成异戊烯基二甲基磷酸酯（IPP; C 5 ）与烯丙基受体，例如二磷酸二甲基烯丙酯（DMAPP; C 5），GPP，FPP或GGPP）由异戊二烯基二磷酸合酶（IDS）催化的反应。用于确定异戊二烯基二磷酸酯产物的IDS酶测定法通常是使用放射性标记的底物进行的，并且所形成的产物是通过使用昂贵的仪器（例如磷光成像仪，radio-GC或radio-HPLC）来鉴定的。尽管已经报道了一种用于测量粗植物提取物中IDS活性的非放射性测定方法，但它需要使用色谱结合串联质谱（LC / MS-MS）的复杂方法。在这里，我们描述了用于确定使用非放射性标记的IPP及其共同烯丙基底物DMAPP，GPP的IDS分析产物非放射性和简单廉价的测定法，和FPP。在测定中生成的异戊二烯基二磷酸产物的检测非常高效，并且在浓度大于40 µM的IPP和DMAPP / GPP / ...
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