Analysis of carbon substrates used by Listeria monocytogenes during growth in J774A.1 macrophages suggests a bipartite intracellular metabolism.
Analysis of carbon substrates used by Listeria monocytogenes during growth in J774A.1 macrophages suggests a bipartite intracellular metabolism.
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对单增李斯特氏菌在 J774A.1 巨噬细胞中生长过程中所使用的碳底物的分析表明，存在双向细胞内代谢。
DOI：
10.3389/fcimb.2014.00156
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发表时间：
2014
影响因子：
5.7
通讯作者：
Eisenreich W
中科院分区：
文献类型：
--
作者：
Grubmüller S;Schauer K;Goebel W;Fuchs TM;Eisenreich W
细胞内细菌病原体（intramellular bacterial pathogens，IBP）依赖于宿主细胞提供的各种营养物质。因此，不同的策略可能是必要的，以适应IBPs的细胞内代谢的宿主细胞。IBPs的具体碳源、参与其降解的分解代谢途径和生物合成性能仍然知之甚少。在这份报告中，我们已经利用了13 C-同位素分析技术，进一步研究单核细胞增生李斯特菌的碳代谢，通过使用EGDe野生型菌株和突变体（在摄取和/或catalysts的各种碳化合物的缺陷）复制在J774A.1巨噬细胞。为此，在[1，2 - 13 C2]葡萄糖、[U-13 C3]甘油、[U-13 C3]丙酮酸、[U-13 C3]乳酸或[U-13 C]氨基酸混合物存在下培养感染的巨噬细胞。基于GC/MS的同位素分析表明，IBP有效利用了氨基酸、葡萄糖6-磷酸、甘油和（在较低程度上）乳酸盐，但不利用丙酮酸盐。大多数从宿主细胞输入的氨基酸直接用于细菌蛋白质的生物合成，几乎不被分解代谢。然而，Asp是由IBP从头合成的，而不是从宿主细胞输入的。正如预期的那样，甘油通过糖酵解途径的ATP生成下部被分解代谢，但显然不用于糖异生。在糖酵解途径的上部和戊糖磷酸分流中由葡萄糖6-磷酸产生的中间体可能主要用于合成代谢目的（可能用于细胞壁组分和核苷酸的生物合成）。这种二分细菌代谢涉及至少两种主要碳底物--主要用于能量供应的甘油和主要用于不可或缺的合成代谢性能的6-磷酸葡萄糖--可能会对受感染的宿主细胞产生更少的营养压力，从而延长宿主细胞的寿命，从而使IBP受益。
Intracellular bacterial pathogens (IBPs) are dependent on various nutrients provided by the host cells. Different strategies may therefore be necessary to adapt the intracellular metabolism of IBPs to the host cells. The specific carbon sources, the catabolic pathways participating in their degradation, and the biosynthetic performances of IBPs are still poorly understood. In this report, we have exploited the technique of 13C-isotopologue profiling to further study the carbon metabolism of Listeria monocytogenes by using the EGDe wild-type strain and mutants (defective in the uptake and/or catabolism of various carbon compounds) replicating in J774A.1 macrophages. For this goal, the infected macrophages were cultivated in the presence of [1,2-13C2]glucose, [U-13C3]glycerol, [U-13C3]pyruvate, [U-13C3]lactate, or a mix of [U-13C]amino acids. GC/MS-based isotopologue profiling showed efficient utilization of amino acids, glucose 6-phosphate, glycerol, and (at a low extent) also of lactate but not of pyruvate by the IBPs. Most amino acids imported from the host cells were directly used for bacterial protein biosynthesis and hardly catabolized. However, Asp was de novo synthesized by the IBPs and not imported from the host cell. As expected, glycerol was catabolized via the ATP-generating lower part of the glycolytic pathway, but apparently not used for gluconeogenesis. The intermediates generated from glucose 6-phosphate in the upper part of the glycolytic pathway and the pentose phosphate shunt likely serve primarily for anabolic purposes (probably for the biosynthesis of cell wall components and nucleotides). This bipartite bacterial metabolism which involves at least two major carbon substrates—glycerol mainly for energy supply and glucose 6-phosphate mainly for indispensible anabolic performances—may put less nutritional stress on the infected host cells, thereby extending the lifespan of the host cells to the benefit of the IBPs.
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影响因子：
3.7
作者：
Gillmaier N;Götz A;Schulz A;Eisenreich W;Goebel W
通讯作者：
Goebel W
影响因子：
3.3
作者：
Donaldson, Janet R.;Nanduri, Bindu;Lawrence, Mark L.
通讯作者：
Lawrence, Mark L.
影响因子：
5.2
作者：
Fuchs TM;Eisenreich W;Kern T;Dandekar T
通讯作者：
Dandekar T
影响因子：
3.6
作者：
Eylert, Eva;Schaer, Jennifer;Eisenreich, Wolfgang
通讯作者：
Eisenreich, Wolfgang
影响因子：
3.1
作者：
Chatterjee, SS;Hossain, H;Hain, T
通讯作者：
Hain, T