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大豆异黄酮代谢体的双锚定:IFS和C4H将复合物拴系在内质网上的证据。

Twin anchors of the soybean isoflavonoid metabolon: evidence for tethering of the complex to the endoplasmic reticulum by IFS and C4H.

作者信息

Dastmalchi Mehran, Bernards Mark A, Dhaubhadel Sangeeta

机构信息

Department of Biology, University of Western Ontario, London, Ontario, Canada.

Agriculture and Agri-Food Canada, 1391 Sandford Street, London, Ontario, Canada.

出版信息

Plant J. 2016 Mar;85(6):689-706. doi: 10.1111/tpj.13137.

Abstract

Isoflavonoids are specialized plant metabolites, almost exclusive to legumes, and their biosynthesis forms a branch of the diverse phenylpropanoid pathway. Plant metabolism may be coordinated at many levels, including formation of protein complexes, or 'metabolons', which represent the molecular level of organization. Here, we have confirmed the existence of the long-postulated isoflavonoid metabolon by identifying elements of the complex, their subcellular localizations and their interactions. Isoflavone synthase (IFS) and cinnamate 4-hydroxylase (C4H) have been shown to be tandem P450 enzymes that are anchored in the ER, interacting with soluble enzymes of the phenylpropanoid and isoflavonoid pathways (chalcone synthase, chalcone reductase and chalcone isomerase). The soluble enzymes of these pathways, whether localized to the cytoplasm or nucleus, are tethered to the ER through interaction with these P450s. The complex is also held together by interactions between the soluble elements. We provide evidence for IFS interaction with upstream and non-consecutive enzymes. The existence of such a protein complex suggests a possible mechanism for flux of metabolites into the isoflavonoid pathway. Further, through interaction studies, we identified several candidates that are associated with GmIFS2, an isoform of IFS, in soybean hairy roots. This list provides additional candidates for various biosynthetic and structural elements that are involved in isoflavonoid production. Our interaction studies provide valuable information about isoform specificity among isoflavonoid enzymes, which may guide future engineering of the pathway in legumes or help overcome bottlenecks in heterologous expression.

摘要

异黄酮是植物特有的代谢产物,几乎仅存在于豆科植物中,其生物合成构成了多样的苯丙烷类途径的一个分支。植物代谢可能在多个层面上进行协调,包括蛋白质复合物或“代谢体”的形成,这代表了分子组织水平。在此,我们通过鉴定该复合物的组成成分、它们的亚细胞定位及其相互作用,证实了长期以来推测存在的异黄酮代谢体的存在。异黄酮合酶(IFS)和肉桂酸4-羟化酶(C4H)已被证明是锚定在内质网中的串联P450酶,它们与苯丙烷类和异黄酮途径的可溶性酶(查尔酮合酶、查尔酮还原酶和查尔酮异构酶)相互作用。这些途径的可溶性酶,无论定位于细胞质还是细胞核,都通过与这些P450的相互作用而与内质网相连。该复合物还通过可溶性成分之间的相互作用而结合在一起。我们提供了IFS与上游和非连续酶相互作用的证据。这种蛋白质复合物的存在表明了代谢产物进入异黄酮途径的一种可能机制。此外,通过相互作用研究,我们在大豆毛状根中鉴定出了几种与IFS的一种同工型GmIFS2相关的候选物。该列表为参与异黄酮生产的各种生物合成和结构成分提供了更多候选物。我们的相互作用研究提供了有关异黄酮酶之间同工型特异性的有价值信息,这可能指导未来豆科植物中该途径的工程改造或有助于克服异源表达中的瓶颈。

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