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牙龈卟啉单胞菌中的新型菌毛蛋白PGN_1808

Novel fimbrilin PGN_1808 in Porphyromonas gingivalis.

作者信息

Nagano Keiji, Hasegawa Yoshiaki, Yoshida Yasuo, Yoshimura Fuminobu

机构信息

Department of Microbiology, School of Dentistry, Aichi Gakuin University 1-100 Kusumoto-cho, Chikusa-ku, Nagoya, Aichi, Japan.

出版信息

PLoS One. 2017 Mar 15;12(3):e0173541. doi: 10.1371/journal.pone.0173541. eCollection 2017.

Abstract

Porphyromonas gingivalis, a periodontopathic gram-negative anaerobic bacterium, generally expresses two types of fimbriae, FimA and Mfa1. However, a novel potential fimbrilin, PGN_1808, in P. gingivalis strain ATCC 33277 was recently identified by an in silico structural homology search. In this study, we experimentally examined whether the protein formed a fimbrial structure. Anion-exchange chromatography showed that the elution peak of the protein was not identical to those of the major fimbrilins of FimA and Mfa1, indicating that PGN_1808 is not a component of these fimbriae. Electrophoretic analyses showed that PGN_1808 formed a polymer, although it was detergent and heat labile compared to FimA and Mfa1. Transmission electron microscopy showed filamentous structures (2‒3 nm × 200‒400 nm) on the cell surfaces of a PGN_1808-overexpressing P. gingivalis mutant (deficient in both FimA and Mfa1 fimbriae) and in the PGN_1808 fraction. PGN_1808 was detected in 81 of 84 wild-type strains of P. gingivalis by western blotting, suggesting that the protein is generally present in P. gingivalis.

摘要

牙龈卟啉单胞菌是一种牙周病相关的革兰氏阴性厌氧菌,通常表达两种菌毛,即FimA和Mfa1。然而,最近通过计算机模拟结构同源性搜索在牙龈卟啉单胞菌ATCC 33277菌株中鉴定出一种新的潜在菌毛蛋白PGN_1808。在本研究中,我们通过实验检测了该蛋白是否形成菌毛结构。阴离子交换色谱显示,该蛋白的洗脱峰与FimA和Mfa1主要菌毛蛋白的洗脱峰不同,这表明PGN_1808不是这些菌毛的组成成分。电泳分析表明,PGN_1808形成了聚合物,尽管与FimA和Mfa1相比,它对去污剂和热不稳定。透射电子显微镜显示,在过表达PGN_1808的牙龈卟啉单胞菌突变体(缺乏FimA和Mfa1菌毛)的细胞表面以及PGN_1808组分中存在丝状结构(2-3nm×200-400nm)。通过蛋白质印迹法在84株牙龈卟啉单胞菌野生型菌株中的81株中检测到了PGN_1808,这表明该蛋白普遍存在于牙龈卟啉单胞菌中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/36ff/5351860/e8c2b83ce947/pone.0173541.g001.jpg

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