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Pathway for lipid A biosynthesis in Arabidopsis thaliana resembling that of Escherichia coli

机译:拟南芥中类似于大肠杆菌的脂质A生物合成途径

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The lipid A moiety of Escherichia coli lipopolysaccharide is a hexa acylated disaccharide of glucosamine that makes up the outer monolayer of the outer membrane. Arabidopsis thaliana contains nuclear genes encoding orthologs of key enzymes of bacterial lipid A biosynthesis, including LpxA, LpxC, LpxD, LpxB, LpxK and KdtA. Although structurally related lipid A molecules are found in most other Gram-negative bacteria, lipid A and its precursors have not been directly detected in plants previously. However, homozygous insertional knockout mutations or RNAi knock-down constructs of Arabidopsis Ipx and kdtA mutants revealed accumulation (or dis appearance) of the expected monosaccharide or disaccharide lipid A precursors by mass spectrometry of total lipids extracted from 10-day old seedlings of these mutants. In addition, fluorescence microscopy of Ipx-gfp fusions in transgenic Arabidopsis plants suggests that the Lpx and KdtA proteins are expressed and tar geted to mitochondria. Although the structure of the lipid A end product generated by plants is still unknown, our work demon strates that plants synthesize lipid A precursors using the same enzymatic pathway present in E. coli.
机译:大肠杆菌脂多糖的脂质A部分是氨基葡萄糖的六酰化二糖,构成外膜的外单层。拟南芥含有编码细菌脂质A生物合成关键酶直系同源基因的核基因,包括LpxA,LpxC,LpxD,LpxB,LpxK和KdtA。尽管在大多数其他革兰氏阴性细菌中都发现了与结构相关的脂质A分子,但以前尚未在植物中直接检测到脂质A及其前体。然而,纯合子插入敲除突变或拟南芥Ipx和kdtA突变体的RNAi敲低构建体通过质谱分析从这些突变体的10天龄幼苗中提取的总脂质,揭示了预期的单糖或二糖脂质A前体的积累(或消失)。 。此外,转基因拟南芥植株中Ipx-gfp融合蛋白的荧光显微镜观察表明,Lpx和KdtA蛋白被表达,焦油到达线粒体。尽管植物产生的脂质A终产物的结构仍是未知的,但我们的工作表明植物利用大肠杆菌中存在的相同酶促途径合成脂质A前体。

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