Purification and characterization of a fimbrial protein from Porphyromonas salivosa ATCC 49407
Purification and characterization of a fimbrial protein from Porphyromonas salivosa ATCC 49407
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DOI：
10.1292/jvms.19-0067
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发表时间：
2019-06-01
影响因子：
1.2
通讯作者：
Hamada, Nobushiro
中科院分区：
文献类型：
--
作者：
Koyata, Yasunori;Watanabe, Kiyoko;Hamada, Nobushiro
牙周病是伴侣动物如狗和猫的一个重要问题。然而，很少有关于伴侣动物的牙周病菌毛协会的信息。在这项研究中，我们已经纯化和其特征在于从唾液卟啉单胞菌ATCC 49407菌毛。该蛋白的分子量约为60 kDa，通过SDS-PAGE估计。免疫金电子显微镜显示，抗60-kDa菌毛血清结合到唾液毕赤酵母ATCC 49407细胞表面的菌毛上。然而，牙龈卟啉单胞菌和牙龈卟啉单胞菌的菌毛没有被相同的抗体标记。免疫电镜研究和免疫印迹分析表明，抗原性和分子量是不同于以前报道的卟啉单胞菌菌毛蛋白。60-kDa的菌毛蛋白的N-末端15个残基的氨基酸序列揭示了15个残基中只有3个与其他卟啉单胞菌属物种的菌毛蛋白相同。因此，唾液毕赤酵母的60-kDa菌毛蛋白的N-末端氨基酸序列明显不同于先前报道的菌毛蛋白。唾液毕赤酵母的粘附率为1.81%。已证实，唾液毕赤酵母可粘附于人体细胞。这些结果表明，60-kDa的菌毛唾液毕赤酵母ATCC 49407是一种新型的菌毛，并可能在粘附宿主细胞的重要因素。我们认为，P. salivosa的表面结构可能在伴侣动物牙周袋中的这种生物体的定植中发挥作用。
Periodontal disease is a significant problem in companion animals such as dogs and cats. However, there is little information available about fimbriae association of periodontal disease in companion animals. In this study, we have purified and characterized a fimbriae from Porphyromonas salivosa ATCC 49407. The molecular mass of this protein was approximately 60-kDa, as estimated by SDS-PAGE. Immunogold electron microscopy revealed that anti-60-kDa fimbrial serum bound to fimbria on the cell surface of P. salivosa ATCC 49407. However, fimbriae of P. gingivalis and P. gulae were not labeled with the same antibody. Immunoelectron-microscopic studies and immunoblot analysis revealed that antigenicity and molecular weight were distinct from previously reported Porphyromonas fimbrial proteins. The amino acid sequence of the N-terminal 15 residues of the 60-kDa fimbrillin protein revealed only 3 of 15 residues identical to other Porphyromonas species fimbrillin proteins. Thus, the N-terminal amino acid sequence of the 60-kDa fimbrillin protein of P. salivosa clearly differed from previously reported fimbrillin proteins. The level of adherence of the P. salivosa was 1.81%. It was confirmed that P. salivosa can adheres to human cells. These results suggest that the 60-kDa fimbriae of P. salivosa ATCC 49407 is a new type of fimbria and may have an important factor in the adherence host cells. We suggest that the surface structure of P. salivosa may have a role in the colonization of this organism in periodontal pockets in companion animals.