Radziejewska-Lebrecht J, Skurnik M, Shashkov A S, Brade L, Rózalski A, Bartodziejska B, Mayer H
Department of Microbiology, University of Silesia, Katowice, Poland.
Acta Biochim Pol. 1998;45(4):1011-9.
Three mutants of Yersinia enterocolitica O:3, namely: YeO3-R1, YeO3-RfbR7 and YeO3-c-trs8-R were classified on the basis of sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS/PAGE) profile of isolated lipopolysaccharides (LPS) as belonging to the Ra- (the first) and the Rc-type (the other two mutants). Methylation analysis, in addition to 13C and 1H NMR studies of purified core oligosaccharides revealed structures similar to those established previously for the full core of Y. enterocolitica O:3 in the case of the Ra mutant, and identical to that reported for the Rc mutant Ye75R, in the case of the two other mutants. The O-specific sugar, 6d-L-altrose, which forms a homopolymeric O-chain, was present in small amounts in all three LPS preparations, as well as in the core oligosaccha ride preparations along with the Ra and the Rc sugars, characteristic of the Y. enterocolitica O:3 core. This result is in line with genetic data, indicating that it is the inner core region which is the receptor for the O-specific chain in Y. enterocolitica O:3. This region seems likewise to be the anchoring region for the enterobacterial common antigen (ECA), as shown by SDS/PAGE/Western blot analysis with monoclonal antibodies against ECA. In addition, we also demonstrated that the Ye75R mutant Rc and its parental strain Ye75S, both were ECA-immunogenic strains. So far, ECA-immunogenic strains, i.e. those with LPS-linked ECA, were only identified in E. coli mutants of the R1, R4 and K-12 serotype.
小肠结肠炎耶尔森氏菌O:3的三个突变体，即YeO3-R1、YeO3-RfbR7和YeO3-c-trs8-R，根据分离的脂多糖（LPS）的十二烷基硫酸钠/聚丙烯酰胺凝胶电泳（SDS/PAGE）图谱，被归类为Ra-型（第一个突变体）和Rc-型（另外两个突变体）。除了对纯化的核心寡糖进行13C和1H NMR研究外，甲基化分析还揭示，在Ra突变体的情况下，其结构与先前确定的小肠结肠炎耶尔森氏菌O:3全核心结构相似；在另外两个突变体的情况下，其结构与报道的Rc突变体Ye75R相同。形成同聚O链的O-特异性糖6d-L-阿洛糖在所有三种LPS制剂中含量较少，在核心寡糖制剂中也与Ra和Rc糖一起存在，这是小肠结肠炎耶尔森氏菌O:3核心的特征。这一结果与遗传数据一致，表明小肠结肠炎耶尔森氏菌O:3中特异性O链的受体是内核区域。如用抗肠杆菌共同抗原（ECA）的单克隆抗体进行SDS/PAGE/蛋白质免疫印迹分析所示，该区域似乎同样是ECA的锚定区域。此外，我们还证明Ye75R突变体Rc及其亲本菌株Ye75S都是ECA免疫原性菌株。到目前为止，ECA免疫原性菌株，即那些LPS连接有ECA的菌株，仅在R1、R4和K-12血清型的大肠杆菌突变体中被鉴定出来。
