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A study Of ora| streptococcal histone-like protein and virulence mechanism

奥拉研究|

基本信息

批准号：
12671827
负责人：
HIROTA Katsuhiko
金额：
$ 2.11万
依托单位：
The University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

Streptococcus intermedius is associated with endogenous infections leading to abscesses in the oral cavity and deep-seated sites, such as the brain and liver. Histone-like protein (HLP), which binds to DNA and RNA molecules and to lipoteichoic acid and epithelial cells, is well conserved among bacterial species and induces macrophages to produce interleukin (IL)-1 and tumor necrosis factor (TNF)-α. In this study, we cloned the gene encoding S. intermedius HLP (HLPSi) from chromosomal DNA by use of the polymerase chain reaction (PCR) using the primers based on S. pyogenes hip gene and ligated to the glutathione S-transferase (GST) fusion vector. The encoded HLPSi had 91 amino acids; and its molecular weight and isoelectric point calculated from the sequence were 9,603 and 10.21 , respectively. The sequence of HLPSi had a high identity with those sequences of the HLP of S. pyogenes A374 and Staphylococcus aureus Mu50 and showed high homology with a 21 -kDa molecule of Mycobavterium leprae (ML-LBP21) at the amino acid level. The immunoelectron micrograph of S. intermedius by anti-HLPSi antibody and ELISA demonstrated that a part of HLPSi was localized on the cell surface and that recombinant HLPSi (rHLPSi) bound to heparin. Our results suggest that HLP produced from bacterial cells may play various roles in pathogenesis by helping the bacteria to bind to their target tissue within the infected host and may contribute to tissue injury at the infection site.

中间链球菌与内源性感染有关，导致口腔和深部部位（如脑和肝）的脓肿。组蛋白样蛋白（HLP）与DNA和RNA分子以及脂磷壁酸和上皮细胞结合，在细菌物种中是很保守的，并诱导巨噬细胞产生白细胞介素（IL）-1和肿瘤坏死因子（TNF）-α。本研究中，我们克隆了S.通过使用基于S. intermedius HLP（HLPSi）的引物的聚合酶链反应（PCR），从染色体DNA中分离HLPSi。化脓性链球菌hip基因，并连接至谷胱甘肽S-转移酶（GST）融合载体。该基因编码的HLPSi由91个氨基酸组成，分子量为9,603，等电点为10.21。HLPSi基因序列与S.化脓性葡萄球菌A374和金黄色葡萄球菌Mu 50，并在氨基酸水平上与麻风分枝杆菌（ML-LBP 21）的21 -kDa分子显示出高度同源性。免疫电镜下观察到S.抗HLPSi抗体和ELISA检测结果表明，HLPSi部分定位于细胞表面，重组HLPSi（rHLPSi）与肝素结合。我们的研究结果表明，从细菌细胞产生的HLP可能发挥各种作用的发病机制，通过帮助细菌结合到它们的目标组织内感染的主机，并可能有助于在感染部位的组织损伤。