Carbon metabolism of Listeria monocytogenes growing inside macrophages
Carbon metabolism of Listeria monocytogenes growing inside macrophages
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DOI：
10.1111/j.1365-2958.2008.06337.x
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发表时间：
2008-08-01
影响因子：
3.6
通讯作者：
Eisenreich, Wolfgang
中科院分区：
文献类型：
--
作者：
Eylert, Eva;Schaer, Jennifer;Eisenreich, Wolfgang
以小鼠J774A.1巨噬细胞为宿主细胞，通过C-13-同位素分析研究了单核细胞增生李斯特菌的细胞内代谢。感染后6小时，将细菌从巨噬细胞中分离并水解。氨基酸转化为叔丁基-二甲基甲硅烷基衍生物，并进行气相色谱/质谱。当巨噬细胞在感染前而不是感染期间被供应[U-C-13（6）]葡萄糖时，标记仅在巨噬细胞的Ala、Asp和Glu中检测到，并且细菌蛋白具有相等的同位素分布。当在感染期间提供[U-C-13（6）]葡萄糖时，在来自宿主和细菌蛋白的Ala、Asp和Glu中以及在来自细菌组分的Ser、Gly、Thr和瓦尔中再次发现C-13标记。L.突变体在相同的实验设置下，在C-3-代谢物甘油和/或二羟基丙酮的摄取和催化方面有缺陷的单核细胞增多症显示出[U-C-13（6）]葡萄糖掺入细菌氨基酸的减少。13 C模式表明：（1）细菌氨基酸的显著部分（50-100%）由宿主细胞提供;（2）C-3代谢产物可作为L.在细胞内条件下的单核细胞增多症和（iii）细菌生物合成的Asp，Thr和Glu的收益通过草酰乙酸通过羧化丙酮酸。
The intracellular metabolism of Listeria monocytogenes was studied by C-13-isotopologue profiling using murine J774A.1 macrophages as host cells. Six hours after infection, bacteria were separated from the macrophages and hydrolyzed. Amino acids were converted into tert-butyl-dimethylsilyl derivatives and subjected to gas chromatography/mass spectrometry. When the macrophages were supplied with [U-C-13(6)]glucose prior to infection, but not during infection, label was detected only in Ala, Asp and Glu of the macrophage and bacterial protein with equal isotope distribution. When [U-C-13(6)]glucose was provided during the infection period, C-13 label was found again in Ala, Asp and Glu from host and bacterial protein, but also in Ser, Gly, Thr and Val from the bacterial fraction. Mutants of L. monocytogenes defective in the uptake and catabolism of the C-3-metabolites, glycerol and/or dihydroxyacetone, showed reduced incorporation of [U-C-13(6)]glucose into bacterial amino acids under the same experimental settings. The 13C pattern suggests that (I) significant fractions (50-100%) of bacterial amino acids were provided by the host cell, (ii) a C-3-metabolite can serve as carbon source for L. monocytogenes under intracellular conditions and (iii) bacterial biosynthesis of Asp, Thr and Glu proceeds via oxaloacetate by carboxylation of pyruvate.