Kogan G, Haraguchi G, Hull S I, Hull R A, Shashkov A S, Jann B, Jann K
Max-Planck-Institut für Immunbiologie, Freiburg, Germany.
Eur J Biochem. 1993 May 15;214(1):259-65. doi: 10.1111/j.1432-1033.1993.tb17919.x.
In previous studies it had been shown that lipopolysaccharide from O4-specific recombinant Escherichia coli, had serological reactivities and a chemical composition that differed from wildtype O4 LPS [Haraguchi, G.E., Zähringer, U., Jann, B., Jann, K., Hull, R.A. & Hull, S.I. (1991) Microb. Pathog. 10, 351-361]. Here we present the structural elucidation of the O-specific moieties from lipopolysaccharides of some of the recombinant strains obtained in previous studies. Compositional analysis, methylation, chemical reactions and NMR spectroscopy showed that, during genetic manipulations (recombination, cosmid cloning, plasmid subcloning), a gradual structural change in the O-specific polysaccharides was observed in the recombinant strains. These changes comprised of an alteration in the position of glucose (side chain) substitution, a change in the anomeric configuration of the main-chain N-acetylglucosamine and an exchange of alpha-L-rhamnopyranose for beta-D-galactofuranose. The relevance of these results for lipopolysaccharide cloning and lipopolysaccharide biosynthesis are discussed.
在先前的研究中已经表明，来自O4特异性重组大肠杆菌的脂多糖具有血清学反应性，并且其化学组成与野生型O4脂多糖不同[原口，G.E.，察林格，U.，扬，B.，扬，K.，赫尔，R.A.和赫尔，S.I.（1991年）《微生物病原体》10，351 - 361]。在此，我们展示了对先前研究中获得的一些重组菌株的脂多糖O特异性部分的结构解析。组成分析、甲基化、化学反应和核磁共振光谱表明，在基因操作（重组、黏粒克隆、质粒亚克隆）过程中，在重组菌株中观察到O特异性多糖的结构逐渐发生变化。这些变化包括葡萄糖（侧链）取代位置的改变、主链N - 乙酰葡糖胺异头构型的变化以及α - L - 鼠李吡喃糖被β - D - 半乳呋喃糖取代。讨论了这些结果对于脂多糖克隆和脂多糖生物合成的相关性。
