Analysis of major virulence factors in Porphyromonas gingivalis under various culture temperatures using specific antibodies
Analysis of major virulence factors in Porphyromonas gingivalis under various culture temperatures using specific antibodies
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DOI：
10.1111/j.1348-0421.2004.tb03552.x
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发表时间：
2004-01-01
影响因子：
2.6
通讯作者：
Yoshimura, F
中科院分区：
文献类型：
--
作者：
Murakami, Y;Masuda, T;Yoshimura, F
牙龈卟啉单胞菌与成人牙周炎的发生有关。我们以前已经确定了主要的外膜蛋白从牙龈卟啉单胞菌，其中包括代表性的毒力因子，如牙龈卟啉单胞菌，一个75 kDa的主要蛋白质，RagA，RagB，和推定的孔蛋白。菌毛是另一种重要的毒力因子，存在于细胞表面。在这项研究中，我们确定了主要的上清液蛋白。它们是75 kDa的主要蛋白质fimbrilin、牙龈卟啉菌蛋白酶及其粘附素结构域。显微镜观察显示，上清蛋白形成囊泡状和菌毛状结构。为了进一步了解该菌的特性，我们研究了生长温度对这些毒力因子的定位和表达的影响。在一般情况下，本地化的主要毒力因子没有改变在不同的生长温度下使用。大多数的75 kDa的主要蛋白质，RagA，RagB，和推定的孔蛋白被发现在包膜部分，而不是在无细胞培养上清液。在包膜部分和上清液中均发现牙龈蛋白酶。80%以上的菌毛与细胞结合，不到20%的菌毛迁移到上清液中。大多数菌毛存在于整个细胞裂解液中，虽然有少量的包膜部分。当生长温度升高时，菌毛、牙龈蛋白酶、75 kDa主要蛋白、RagA和RagB的表达降低。然而，温度几乎没有影响的推定孔蛋白的表达。主要毒力因子的表达在较高温度下降低的趋势可能使牙龈卟啉单胞菌能够在恶劣条件下生存。
Porphyromonas gingivalis is implicated in the occurrence of adult periodontitis. We have previously identified major outer membrane proteins from R gingivalis, which include representative virulence factors such as gingipains, a 75 kDa major protein, RagA, RagB, and putative porin. Fimbriae, another important virulence factor, exist on the cell surface. In this study, we identified major supernatant proteins. They were fimbrilin, the 75 kDa major protein, gingipains and their adhesin domains. Microscopic examination showed that supernatant proteins formed vesicle-like and fimbrial structures. To learn more about the character of this bacterium, we examined effects of growth temperature on localization and expression of these virulence factors. In general, localization of major virulence factors did not change at the various growth temperatures used. Most of the 75 kDa major protein, RagA, RagB, and putative porin were found in the envelope fraction, not in cell-free culture supernatant. Gingipains; were found in both the envelope fraction and supernatant. More than 80% of fimbriae were associated with cells, less than 20% migrated to the supernatant. Most fimbriae existed in the whole cell lysate, although there was a small amount in the envelope fraction. When the growth temperature was increased, expression of fimbriae, gingipains, the 75 kDa major protein, RagA, and RagB decreased. However, temperature had almost no effect on expression of putative porin. The tendency for expression of major virulence factors to decrease at higher temperatures may enable P gingivalis to survive under hostile conditions.