Imai Masashi, Murakami Yukitaka, Nagano Keiji, Nakamura Hiroshi, Yoshimura Fuminobu
Department of Microbiology, School of Dentistry, Aichi-Gakuin University, 1-100 Kusumoto-cho, Chikusa-ku, Nagoya, Aichi, Japan.
Eur J Oral Sci. 2005 Oct;113(5):391-9. doi: 10.1111/j.1600-0722.2005.00235.x.
Porphyromonas gingivalis has been implicated in both marginal periodontitis and periapical infection. This study examined the major outer membrane proteins, from P. gingivalis, which related to periradicular lesions. Outer membrane protein profiles of P. gingivalis ATCC 33277 and W83 were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and N-terminal amino acid analysis. Most outer membrane proteins, such as RagA, gingipains, and OmpA-like proteins, were found in both strains in a similar distribution pattern; however, the migration positions of Lys-gingipain and RagB were inverted in SDS-PAGE. Western blot analysis showed that RagA, RagB, and OmpA-like proteins were found in all of the P. gingivalis strains tested. The antiserum of W83 against RagB reacted poorly to some strains, such as ATCC 33277. When strains phylogenetically related to P. gingivalis were examined, RagA and OmpA homologs were immunologically detected in several strains. However, none of the RagB homologs were detected in any strain analyzed, suggesting that RagB is unique to P. gingivalis. To examine immunoreactive antigens in P. gingivalis, sera from patients with periradicular lesions were used. More than half of the sera showed strong reactions to P. gingivalis cell components, especially RagB. Our results indicate that a major outer membrane protein, RagB, is a possible virulence factor in periradicular lesions.
牙龈卟啉单胞菌与边缘性牙周炎和根尖周感染均有关联。本研究检测了牙龈卟啉单胞菌中与根尖周病变相关的主要外膜蛋白。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和N端氨基酸分析,比较了牙龈卟啉单胞菌ATCC 33277和W83的外膜蛋白谱。大多数外膜蛋白,如RagA、牙龈蛋白酶和OmpA样蛋白,在两种菌株中的分布模式相似;然而,赖氨酸牙龈蛋白酶和RagB在SDS-PAGE中的迁移位置相反。蛋白质印迹分析表明,在所有测试的牙龈卟啉单胞菌菌株中均发现了RagA、RagB和OmpA样蛋白。W83针对RagB的抗血清与某些菌株(如ATCC 33277)反应较弱。当检测与牙龈卟啉单胞菌系统发育相关的菌株时,在几种菌株中免疫检测到了RagA和OmpA同源物。然而,在任何分析的菌株中均未检测到RagB同源物,这表明RagB是牙龈卟啉单胞菌特有的。为了检测牙龈卟啉单胞菌中的免疫反应性抗原,使用了根尖周病变患者的血清。超过一半的血清对牙龈卟啉单胞菌细胞成分有强烈反应,尤其是RagB。我们的结果表明,一种主要的外膜蛋白RagB可能是根尖周病变中的一种毒力因子。