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Novel Modification of Lipid A of Francisella tularensis

机译：图拉弗朗西斯菌脂质A的新修饰

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We have investigated the lipid A of Francisella tularensis subsp. holarctica strain 1547-57, a type B strain, by using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, nanoelectrospray quadrupole ion-trap mass spectrometry, and chemical methods. In accordance with the previously published structures of the lipid A from F. tularensis live vaccine strain (LVS) (ATCC 29684) (E. Vinogradov et al., Eur. J. Biochem. >269:6112-6118, 2002), all of the major lipid A forms from strain 1547-57 were tetraacylated. As in the LVS strain, the major fatty acids detected in the F. tularensis 1547-57 lipid A sample included 3-hydroxyoctadecanoic acid, 3-hydroxyhexadecanoic acid, hexadecanoic acid, and tetradecanoic acid. However, several of the lipid A components present in strain 1547-57 were of higher molecular weight than the previously published structures. A major component with an Mr of 1,666 was found to contain three C18:0(3-OH) fatty acids, one C16:0 fatty acid, one phosphate group, and one 161-Da moiety. This 161-Da moiety could be removed from the lipid A by treatment with aqueous hydrofluoric acid and was identified as galactosamine following peracetylation and analysis by gas chromatography-mass spectrometry. Detailed investigations of the Mr-1,666 species by ion-trap mass spectrometry with multiple stages of fragmentation suggested that the galactosamine-1-phosphate was linked to the reducing terminus of the lipid A. Similar to the modification of lipid A with arabinosamine, lipopolysaccharide species from F. tularensis containing a phosphate-linked galactosamine could potentially influence its intracellular survival by conferring resistance to antimicrobial peptides.
机译：我们已经研究了杜拉弗朗西斯菌亚种的脂质A。通过使用基质辅助激光解吸电离飞行时间质谱法，纳米电喷雾四极杆离子阱质谱法和化学方法对B株 holarctica 株1547-57进行分析。根据先前公开的来自 F的脂质A的结构。 tularensis活疫苗株（LVS）（ATCC 29684）（E。Vinogradov等人，Eur。J. Biochem。> 269： 6112-6118，2002），所有主要脂质将来自菌株1547-57的形式四酰化。与LVS菌株一样，在 F中检测到主要脂肪酸。 tularensis 1547-57脂质样品包括3-羟基十八烷酸，3-羟基十六烷酸，十六烷酸和十四烷酸。但是，菌株1547-57中存在的几种脂质A组分的分子量均高于先前公布的结构。发现 M r 为1,666的主要成分包含三种C 18：0 （3-OH）脂肪酸，一种C < sub> 16：0 脂肪酸，一个磷酸基和一个161-Da部分。可以通过用氢氟酸水溶液处理从脂质A中除去该161-Da部分，并在过乙酰化和通过气相色谱-质谱分析后鉴定为半乳糖胺。离子阱质谱法对 M r -1,666种物种进行了详细的研究，发现该化合物具有多个片段化阶段，表明半乳糖胺-1-磷酸与该蛋白的还原末端相连。脂质A。类似于用阿拉伯糖胺对脂质A的修饰，来自 F的脂多糖种类。含有磷酸酯连接的半乳​​糖胺的土拉菌可能通过赋予其对抗菌肽的抗性而潜在地影响其细胞内存活。

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