Pei Z, Burucoa C, Grignon B, Baqar S, Huang X Z, Kopecko D J, Bourgeois A L, Fauchere J L, Blaser M J
Department of Medicine, Vanderbilt University School of Medicine and Veterans Affairs Medical Center, Nashville, Tennessee 37232, USA.
Infect Immun. 1998 Mar;66(3):938-43. doi: 10.1128/IAI.66.3.938-943.1998.
Campylobacter jejuni is one of the leading causes of bacterial diarrhea throughout the world. We previously found that PEB1 is a homolog of cluster 3 binding proteins of bacterial ABC transporters and that a C. jejuni adhesin, cell-binding factor 1 (CBF1), if not identical to, contains PEB1. A single protein migrating at approximately 27 to 28 kDa was recognized by anti-CBF1 and anti-PEB1. To determine the role that the operon encoding PEB1 plays in C. jejuni adherence, peb1A, the gene encoding PEB1, was disrupted in strain 81-176 by insertion of a kanamycin resistance gene through homologous recombination. Inactivation of this operon completely abolished expression of CBF1, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. In comparison to the wild-type strain, the mutant strain showed 50- to 100-fold less adherence to and 15-fold less invasion of epithelial cells in culture. Mouse challenge studies showed that the rate and duration of intestinal colonization by the mutant were significantly lower and shorter than with the wild-type strain. In summary, PEB1 is identical to a previously identified cell-binding factor, CBF1, in C. jejuni, and the peb1A locus plays an important role in epithelial cell interactions and in intestinal colonization in a mouse model.
空肠弯曲菌是全球细菌性腹泻的主要病因之一。我们之前发现PEB1是细菌ABC转运蛋白3型结合蛋白的同源物,并且空肠弯曲菌的一种粘附素,细胞结合因子1(CBF1),即便不完全相同,也含有PEB1。一种分子量约为27至28 kDa的单一蛋白质可被抗CBF1和抗PEB1识别。为了确定编码PEB1的操纵子在空肠弯曲菌黏附中所起的作用,通过同源重组插入卡那霉素抗性基因,在81-176菌株中破坏了编码PEB1的基因peb1A。如通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹所确定的,该操纵子的失活完全消除了CBF1的表达。与野生型菌株相比,突变菌株在培养物中对上皮细胞的黏附减少了50至100倍,侵袭减少了15倍。小鼠攻毒研究表明,突变体在肠道定殖的速率和持续时间明显低于野生型菌株且更短。总之,在空肠弯曲菌中,PEB1与先前鉴定的细胞结合因子CBF1相同,并且peb1A基因座在小鼠模型的上皮细胞相互作用和肠道定殖中起重要作用。