Antigenic group II chaperonin in Methanobrevibacter oralis may cross-react with human chaperonin CCT.
Antigenic group II chaperonin in Methanobrevibacter oralis may cross-react with human chaperonin CCT.
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DOI：
10.1111/j.2041-1014.2009.00548.x
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发表时间：
2010-04
影响因子：
3.7
通讯作者：
Kokoro Yamabe;H. Maeda;S. Kokeguchi;Y. Soga;M. Meguro;K. Naruishi;S. Asakawa;S. Takashiba
中科院分区：
文献类型：
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作者：
Kokoro Yamabe;H. Maeda;S. Kokeguchi;Y. Soga;M. Meguro;K. Naruishi;S. Asakawa;S. Takashiba
口腔甲烷预防菌是一种经常从严重牙周炎部位分离出来的古细菌。然而，其致病作用尚不清楚。在此，我们旨在从口腔分枝杆菌中分离II族伴侣蛋白并检测其抗原性。利用聚合酶链反应和基因组行走技术，从口腔分枝杆菌中克隆到了两个伴侣蛋白亚基（Cpn-1和Cpn-2）的编码基因。生成重组蛋白Cpn-1和Cpn-2，用Western免疫印迹法检测牙周炎患者血清的反应性。Cpn-1和Cpn-2基因的开放阅读框分别由1641和1614个碱基对组成。在这两个基因中都观察到伴侣蛋白家族中保守的假定的atp结合域。两个基因的氨基酸序列与人类CCT亚基（CCT1-8）的同源性为28.8 ~ 40.0%。36例患者血清中分别有30例和29例与重组Cpn-1和重组Cpn-2反应。使用抗人CCT亚基血清进行免疫印迹检测表明，抗cct3和抗cct8抗体能够识别重组Cpn-1。此外，抗cct1、CCT3、CCT6和CCT8抗体在口腔分枝杆菌中识别约60 kDa的抗原。结果提示口腔分枝杆菌的伴侣蛋白亚基是被牙周炎患者识别的抗原分子，可能与人伴侣蛋白CCT发生交叉反应。
Methanobrevibacter oralis is an archaeal species frequently isolated from sites of severe periodontitis. However, its pathogenic roles remain unclear. Here, we aimed to isolate group II chaperonin from M. oralis and examine its antigenicity. The genes encoding two chaperonin subunits (Cpn-1 and Cpn-2) were cloned from M. oralis using polymerase chain reaction and genome walking procedures. Recombinant proteins Cpn-1 and Cpn-2 were generated, and the reactivities of sera from patients with periodontitis were examined by Western immunoblotting. The open reading frames of Cpn-1 and Cpn-2 genes consisted of 1641 and 1614 base pairs, respectively. Putative ATP-binding domains conserved among the chaperonin family were observed in both genes. The deduced amino acid sequences of the two genes showed 28.8-40.0% identity to each of the subunits of human CCT (CCT1-8). Thirty and 29 of 36 patients' sera reacted with the recombinant Cpn-1 and recombinant Cpn-2, respectively. Western immunoblotting using antiserum against human CCT subunits indicated that anti-CCT3 and anti-CCT8 antibodies recognized recombinant Cpn-1. In addition, anti-CCT1, CCT3, CCT6, and CCT8 antibodies recognized an antigen of approximately 60 kDa in M. oralis. The results suggested that the chaperonin subunits of M. oralis were antigenic molecules that were recognized by periodontitis patients and that may cross-react with human chaperonin CCT.