ANALYSIS OF LIPOPOLYSACCHARIDE BIOSYNTHESIS IN SALMONELLA-TYPHIMURIUM AND ESCHERICHIA-COLI BY USING AGENTS WHICH SPECIFICALLY BLOCK INCORPORATION OF 3-DEOXY-D-MANNO-OCTULOSONATE
ANALYSIS OF LIPOPOLYSACCHARIDE BIOSYNTHESIS IN SALMONELLA-TYPHIMURIUM AND ESCHERICHIA-COLI BY USING AGENTS WHICH SPECIFICALLY BLOCK INCORPORATION OF 3-DEOXY-D-MANNO-OCTULOSONATE
复制标题
DOI：
10.1128/jb.170.5.2185-2191.1988
复制
发表时间：
1988-05-01
影响因子：
3.2
通讯作者：
CAPOBIANCO, JO
中科院分区：
文献类型：
--
作者：
GOLDMAN, RC;DORAN, CC;CAPOBIANCO, JO
使用特异性抑制CTP：CMP-3-脱氧-D-甘露-辛酮糖酸胞苷酰转移酶活性的抗菌剂来阻断3-脱氧-D-甘露-辛酮糖酸（KDO）掺入脂多糖中。脂多糖合成停止，分子类似于脂质A积累的结构，并停止细菌生长后，添加这些代理商培养的鼠伤寒沙门氏菌和大肠杆菌。虽然四种主要的脂质A在S.鼠伤寒沙门氏菌，它们的积累动力学不同。主要种类中极性最小的是IVA [O-（2-氨基-2-脱氧-β- D-吡喃葡萄糖基）-（1 → 6）-2-氨基-2-脱氧-α- D-葡萄糖，在位置2、3、2“和3”用β-葡萄糖酰化羟基肉豆蔻酰基并且在位置1和4“处带有磷酸盐]，一种先前从含有kdsA突变的细菌中分离的分子（C. R. H. Raetz，S.珀塞尔，M. V. Meyer，N. Qureshi和K. Takayama，J.Biol.Chem.260：16080-16088，1985）。种IVA积累的第一和最大程度的抑制剂后，与其他更极性的衍生物出现后，IVA达到一半的最大水平。相比之下，只有两种主要的前体积累在E。在添加抑制剂后，其中一个种与A.根据化学组成、快原子轰击质谱和硅胶H上的共迁移，鼠伤寒沙门氏菌是一种具有相关结构的更极性的物种，并且它也先于更极性的物种积累。我们的结论是，KDO加入到前体物质IVA是脂质A-KDO形成的主要途径。typhimurium LT 2和E.大肠杆菌中，并且缺乏KDO的极性更强的物质的积累仅响应于正常途径抑制后物质IVA的积累而发生。
Antibacterial agents which specifically inhibit CTP:CMP-3-deoxy-D-manno-octulosonate cytidylyltransferase activity were used to block the incorporation of 3-deoxy-D-manno-octulosonate (KDO) into lipopolysaccharide. Lipopolysaccharide synthesis ceased, molecules similar in structure to lipid A accumulated, and bacterial growth ceased following addition of such agents to cultures of Salmonella typhimurium and Escherichia coli. Although four major species of lipid A accumulated in S. typhimurium, their kinetics of accumulation were different. The least polar of the major species was IVA [O-(2-amino-2-deoxy-.beta.-D-glucopyranosyl)-(1.fwdarw.6)-2-amino-2-deoxy-.alpha.-D-glucose, acylated at positions 2, 3, 2'', and 3'' with .beta.-hydroxymyristoyl groups and bearings phosphates at positions 1 and 4''], a molecule previously isolated from bacteria containing a kdsA mutation (C. R. H. Raetz, S. Purcell, M. V. Meyer, N. Qureshi, and K. Takayama, J. Biol. Chem. 260:16080-16088, 1985). Species IVA accumulated first and to the greatest extent following addition of the inhibitor, with other more polar derivatives appearing only after IVA attained half its maximal level. In contrast, only two major species of precursor accumulated in E. coli following addition of the inhibitor. One of these species was identical to IVA from A. typhimurium on the basis of chemical composition, fast atom bombardment mass spectroscopy, and comigration on Silica Gel H, and it also accumulated prior to a more polar species of related structure. We conclude that the addition of KDO to precursor species IVA is the major pathway of lipid A-KDO formation in both S. typhimurium LT2 and E. coli and that accumulation of the more polar species lacking KDO only occurs in response to accumulation of species IVA following inhibition of the normal pathway.