Szymanski C M, Yao R, Ewing C P, Trust T J, Guerry P
Enteric Diseases Program, Naval Medical Research Center, Rockville, MD, USA.
Mol Microbiol. 1999 Jun;32(5):1022-30. doi: 10.1046/j.1365-2958.1999.01415.x.
A genetic locus from Campylobacter jejuni 81-176 (O:23, 36) has been characterized that appears to be involved in glycosylation of multiple proteins, including flagellin. The lipopolysaccharide (LPS) core of Escherichia coli DH5alpha containing some of these genes is modified such that it becomes immunoreactive with O:23 and O:36 antisera and loses reactivity with the lectin wheat germ agglutinin (WGA). Site-specific mutation of one of these genes in the E. coli host causes loss of O:23 and O:36 antibody reactivity and restores reactivity with WGA. However, site-specific mutation of each of the seven genes in 81-176 failed to show any detectable changes in LPS. Multiple proteins from various cellular fractions of each mutant showed altered reactivity by Western blot analyses using O:23 and O:36 antisera. The changes in protein antigenicity could be restored in one of the mutants by the presence of the corresponding wild-type allele in trans on a shuttle vector. Flagellin, which is known to be a glycoprotein, was one of the proteins that showed altered reactivity with O:23 and O:36 antiserum in the mutants. Chemical deglycosylation of protein fractions from the 81-176 wild type suggests that the other proteins with altered antigenicity in the mutants are also glycosylated.
空肠弯曲菌81 - 176(O:23, 36)的一个基因位点已被鉴定,该位点似乎参与了多种蛋白质的糖基化,包括鞭毛蛋白。含有其中一些基因的大肠杆菌DH5α的脂多糖(LPS)核心被修饰,使其与O:23和O:36抗血清具有免疫反应性,并失去与凝集素麦胚凝集素(WGA)的反应性。在大肠杆菌宿主中对这些基因之一进行位点特异性突变会导致O:23和O:36抗体反应性丧失,并恢复与WGA的反应性。然而,对81 - 176中七个基因中的每一个进行位点特异性突变均未显示LPS有任何可检测到的变化。使用O:23和O:36抗血清通过蛋白质印迹分析,每个突变体不同细胞组分中的多种蛋白质显示出反应性改变。在一个突变体中,通过在穿梭载体上反式存在相应的野生型等位基因,可以恢复蛋白质抗原性的变化。已知作为糖蛋白的鞭毛蛋白是在突变体中与O:23和O:36抗血清反应性改变的蛋白质之一。对81 - 176野生型蛋白质组分进行化学去糖基化表明,突变体中其他抗原性改变的蛋白质也被糖基化。