pckA-deficient Porphyromonas gingivalis W83 shows reduction in hemagglutination activity and alteration in the distribution of gingipain activity
pckA-deficient Porphyromonas gingivalis W83 shows reduction in hemagglutination activity and alteration in the distribution of gingipain activity
复制标题
pckA 缺陷的牙龈卟啉单胞菌 W83 显示血凝活性降低和牙龈蛋白酶活性分布改变
DOI：
10.1111/eos.12565
复制
发表时间：
2018
影响因子：
1.9
通讯作者：
Wu Yafei
中科院分区：
文献类型：
--
作者：
Wu Leng;Zhao Lei;Wang Jun;Liu Chengcheng;Li Yan;Wu Yafei
细菌在感染过程中的代谢与细菌的持久性和毒力因素有关。牙龈卟啉单胞菌是导致慢性牙周炎的关键病原体。我们先前的研究表明，编码磷酸烯醇式丙酮酸羧基激酶的基因pck A可能是OFP强毒株的特异性致病基因。牙周炎。在此，构建了APKKA缺陷株(ΔpG1676)。牙周炎W83。对突变株和野生型菌株的毒力特性进行了比较。具体来说，血凝活性是通过凝集绵羊红细胞的能力来确定的。使用合成的特定底物检测姜辣素的活性。用RT-qPCR分析基因表达水平，用银染和电子显微镜检测细胞表面相关多糖。与野生型菌株相比，pck A基因的失活不影响细菌的生长和内毒素的形成，但导致血凝活性降低和血凝相关基因RFA的表达下调。此外，ΔpG1676突变体的牙周疼痛活性分布也发生了改变。在细胞表面检测到牙周疼痛活性增加，但在培养上清液中其活性降低。综上所述，我们的结果表明，pck A基因在调节毒力OFP方面发挥了作用。牙周炎W83。
Bacterial metabolism during infection is related to bacterial persistence and virulence factors.Porphyromonas gingivalisis a key pathogen that contributes to chronic periodontitis. Our previous study showed thatpckA, the gene encoding phosphoenolpyruvate carboxykinase, is a putative‐specific pathogenic gene of virulent strains ofP. gingivalis. Here, apckA‐deficient strain (ΔPG1676) was constructed inP. gingivalisW83. Virulence properties were compared between the mutant and wild‐type strains. Specifically, hemagglutination activity was determined by the ability to agglutinate sheep erythrocytes. Gingipain activity was detected using synthetic‐specific substrates. Gene expression levels were analyzed using RT‐qPCR, and cell surface‐associated polysaccharides were examined by silver staining and electron microscopy. Inactivation of thepckAgene did not affect bacterial growth and lipopolysaccharide formation but led to a reduction in hemagglutination activity and downregulation in expression of the hemagglutination‐associated gene,rfa, when compared with the wild‐type strain. Additionally, the ΔPG1676 mutant exhibited an alteration in the distribution of gingipain activity. Increased gingipain activity was detected on the cell surface, but a decrease in its activity in the culture supernatant was shown. Taken together, our results suggest that thepckAgene plays a role in modulating the virulence ofP. gingivalisW83.