Evidence for a system of general protein glycosylation in Campylobacter jejuni
Evidence for a system of general protein glycosylation in Campylobacter jejuni
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DOI：
10.1046/j.1365-2958.1999.01415.x
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发表时间：
1999-06-01
影响因子：
3.6
通讯作者：
Guerry, P
中科院分区：
文献类型：
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作者：
Szymanski, CM;Yao, RJ;Guerry, P
已对来自空肠弯曲杆菌81-176（O：23，36）的遗传位点进行了表征，该基因座似乎参与多种蛋白质（包括鞭毛蛋白）的糖基化。含有这些基因中的一些的大肠杆菌DH 5 α的脂多糖（LPS）戈尔被修饰，使得其变得与0：23和0：36抗血清具有免疫反应性，并且失去与凝集素麦胚凝集素（WGA）的反应性。在E.大肠杆菌宿主导致0：23和0：36抗体反应性丧失，并恢复与WGA的反应性。然而，在81-176中的7个基因中的每一个的位点特异性突变未能显示LPS中的任何可检测的变化。从每个突变体的各种细胞组分的多种蛋白质显示出改变的反应性通过Western印迹分析使用0：23和0：36抗血清。蛋白质抗原性的变化可以恢复在一个突变体的存在下，相应的野生型等位基因的反式穿梭载体。鞭毛蛋白是已知的糖蛋白，是在突变体中与0：23和0：36抗血清显示出改变的反应性的蛋白质之一。来自81-176野生型的蛋白质级分的化学去糖基化表明突变体中具有改变的抗原性的其它蛋白质也被糖基化。
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) Gore of Escherichia coli DH5 alpha containing some of these genes is modified such that it becomes Immunoreactive with 0:23 and 0:36 antisera and loses reactivity with the lectin wheat germ agglutinin (WGA). She-specific mutation of one of these genes in the E. coli host causes loss of 0:23 and 0:36 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 0:23 and 0:36 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 0:23 and 0:36 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 glycosyrated.